ExperimentCapillaryELISA
ExperimentCapillaryELISA[Samples]⟹Protocol
creates a Protocol to run capillary Enzyme-Linked Immunosorbent Assay (ELISA) experiment on the provided Samples for the detection of certain analytes.
Capillary Enzyme-Linked Immunosorbent Assay (ELISA) uses pre-kitted microfluidic cartridge plates to provide the entire workflow for sandwich ELISA tests. Each microfluidic channel in capillary ELISA cartridge contains multiple Glass Nano-Reactors (GNR) that are functionalized with immunoassay reagents. After loading into the cartridge, samples and reagents are directed into and out of the microfluidics channels through a series of pneumatic pistons. When the samples run through the microfluidic channels, analytes (e.g., peptides, proteins, antibodies and hormones) of interest bind to the immobilized capture antibodies inside GNR through antigen-antibody interaction. Wash buffer is then used to flush any unbound analytes in the microfluidic channels into the waste collection well. In the last step, the detection reagents are added into the microfluodic channels to provide detectable fluorescence signals through their interaction with immobilized analytes. Fluorescence signals are detected with a CCD camera through the transparent GNR. Overall, ExperimentCapillaryELISA is designed for accurate measurements of analyte concentrations in the samples using built-in or customized standard curves, with internal replicates provided.
Experimental Principles
Figure 1.1: Procedural overview of a capillary Enzyme-Linked Immunosorbent Assay (ELISA) experiment. Step 1: Samples are mixed with SpikeSample (optional) and then diluted (mandatory). Dilution is required on all the loading samples for the best performance of microfludics. Spike samples containing known concentration(s) of analyte(s) are used to increase the analytical response of the unknown samples to evaluate the sample matrices in a new ELISA assay. Standard samples are resuspended into solution phase (if originally in solid state) and then diluted. For a customizable capillary ELISA cartridge, capture antibody samples and detection atnibody samples are resuspended into solution phase (if originally in solid state), conjugated with digoxigenin and biotin, respectively, and then diluted to the desired concentrations. Samples, standard samples and antibodies of desired concentrations are loaded into different wells into the capillary ELISA cartridge using a liquid handler. Step 2: The sandwich ELISA process happens inside the Glass Nano-Reactors (GNR). The flow of reagents is precisely controlled by the solenoid valves and the pneumatic pump to allow the sufficient reaction of each step in the entire workflow of the sandwich ELISA experiments inside the microfluidic channels. Step 3: In a customizable cartridge, the capture antibodies are immobilized to the GNR through their specific interactions with the anti-digoxigenin antibody coating. Cartridges with pre-loaded capture antibodes are available for validated ELISA assays through the instrument manufacturer. Analytes of interest in the sample bind to the immobilized capture antibodies inside GNR through antigen-antibody interaction. The detection antibodies bind to analyte epitopes distinct from the capture antibody through antigen-antibody interaction. The detection antibodies bind to analyte epitopes distinct from the capture antibodies through antigen-antibody interaction. Through the biotin-streptavidin interaction, fluorescent labels are added to interact with detection antibodies to provide detectable fluorescence signals. Step 4: Detectors inside the capillary ELISA instrument scan the fluorescent signals inside the transparent GNRs. For a pre-loaded capillary ELISA cartridge, the instrument uses the standard curves provided by the assay manufacturer to calculate the concentrations of analytes in the SamplesIn.
Instrumentation
Ella
Figure 2.1.1: Capillary ELISA instrument (Protein Simple Ella) uses a pneumatic pump to control the flow of reagents and to achieve the entire workflow of sandwich ELISA in the capillary ELISA cartridge at 35 Celsius. The pneumatic pump provides unregulated vacuum and regulated pressure to a manifold upon which 7 solenoid valves (vaccum and pressure selected) are mounted. The cartridge then interfaces with the output of those 7 pneumatic lines to control valves and pistons in the microfluidic channels. The cartridge has a pneumatic section and a fluidic section that are separated with a thin rubber membrane. Pressure from the pneumatic pump through a certain pneumatic line can push the membrane into the fluid and close a valve. On the other hand, vaccum from the pneumatic pump through a certain pneumatic line can pull the fluid and the membrane to open a valve. By opening the specific valves in the capillary ELISA cartridge, the instrument can load capture antibody, sample, wash buffer, detection antibody and fluorescent dye into the microfluidic channels sequentially while collecting the excess reagents into the designated waste well. In each step, through the precise control of valves, the reagents flow through the Glass Nano-Reactors (GNR) multiple times to allow sufficient reactions. After the completion of the entire sandwich ELISA workflow, the instrument acts as a standard epifluorescent system. The fluorescent dye molecules are excited by a power and temperature controlled 25 Milliwatt single mode 641 nm laser diode. The 650 nm fluorescent signal passes through a dichroic bandpass filter and the intensity is measured by a monochrome 12 bit CCD camera with auto-exposure ranging from 10 Microsecond to 15 Millisecond. The background signal for fluorescence measurement is acquired at the positions without Glass Nano-Reactors (GNR) along the microfluidic channels. At each GNR, the fluorescence signal is read 50 times and the mean value of all readings is reported for the sample. The process is identical for each analyte and each sample in the capillary ELISA cartridge.
Experiment Options
General
Instrument
The detection and quantification capillary ELISA device on which the protocol is to be run. The instrument accepts the cartridge loaded with samples and buffers, loads the samples into the capillaries, performs the ELISA experiment and detects the signals to quantify certain analytes.
Pattern Description: An object of type or subtype Model[Instrument, CapillaryELISA] or Object[Instrument, CapillaryELISA]
Programmatic Pattern: ObjectP[{Model[Instrument, CapillaryELISA], Object[Instrument, CapillaryELISA]}]
CartridgeType
The type of capillary ELISA cartridge (SinglePlex72X1, MultiAnalyte32X4, MultiAnalyte16X4, MultiPlex32X8 or Customizable) that is used in the capillary ELISA experiment. For pre-loaded cartridge types, different cartridge types provide different numbers of capacity and throughput (Figure 3.1 in ExperimentCapillaryELISA help file).
Default Calculation: CartridgeType is automatically selected based on the total number of samples (with standard samples and dilutions considered) and the Cartridge option. When the total number of samples is below or equal to 48 and Cartridge option is not Null, CartridgeType is set to Customizable. Otherwise it is set to SinglePlex72X1.
Pattern Description: SinglePlex72X1, MultiAnalyte32X4, MultiAnalyte16X4, MultiPlex32X8, or Customizable.
Cartridge
The single-use capillary ELISA cartridge plate that is used in the instrument to provide the entire workflow of a sandwich ELISA assay and quantify the analytes (such as peptides, proteins, antibodies and hormones) in the samples. A customizable capillary ELISA cartridge Model[Container,Plate,Irregular,CapillaryELISA,"Human 48-Digoxigenin Cartridge"] uses an anti-digoxigenin antibody coating in GNR to serve as the foundation. Digoxigenin-modified capture antibody sample and biotinylated detection antibody sample are prepared prior to the experiment and loaded into the Cartridge for sandwich ELISA experiment of up to 48 samples. For pre-loaded cartridge types (all types except Customizable), the cartridge is loaded with capture antibody and detection antibody reagents, meaning that Cartridge option also determines the ELISA assay to be run and the analytes to be detected in the experiment. For the current list of available capillary ELISA cartridges, please perform Search[Model[Container,Plate,Irregular,CapillaryELISA]] with the desired CartridgeType, AnalyteMolecules or other conditions. If a new model of pre-loaded cartridge is required for the experiment, please set Cartridge option to Null and input more information in CartridgeType, Species and Analytes options. The new model is created and the cartridge is fulfilled accordingly. For a complete list of available analytes on pre-loaded cartridges and their ordering information, please refer to Object[Report,Literature,"Simple Plex Assay Menu"] and Object[ManufacturingSpecification,CapillaryELISACartridge] of different analytes for more details.
Figure 3.1: There are five types of capillary ELISA cartridges. The presented diagram shows a miniaturization of the microfluidic channel system for a single sample with the reagents that should be loaded into the cartridge marked. A customizable cartridge can be loaded with the user's antibody reagents and used for customized sandwich ELISA experiments. One analyte can be tested per sample in one experiment. A customizable capillary ELISA cartridge uses an anti-digoxigenin antibody coating in the Glass Nano-Reactors (GNR) to serve as the foundation for the assay and uses streptavidin-conjugated fluorophore dye reagent for signal detection. The digoxigenin-conjugated capture antibody samples and biotin-conjugated detection antibody samples can be supplied in ExperimentCapillaryELISA or prepared using the corresponding antibody conjugation options. Other antibody preparation options can be specified to achieve the desired antibody resuspension and/or dilution. Capture antibody samples are conjugated with digoxigenin through the reactions between digoxigenin NHS-ester reagents and the primary amines of the antibody samples. Then the conjugated capture antibody samples can be immobilized to the GNR for analytes binding. Detection antibody samples are conjugated with biotin through the reactions between Biotin-XX-SE (6-((6-((Biotinoyl)Amino)Hexanoyl)Amino)Hexanoic Acid, Succinimidyl Ester) and the primary amines of the antibodies, which allows their specific interaction with streptavidin-conjugated fluorophore reagents to provide detectable fluorescent signals. Please refer to Object[Report,Literature," CapillaryELISA 48-Digoxigenin Cartridge Guide "] for details. The other cartridge types - SinglePlex72X1, MultiAnalyte32X4, MultiAnalyte16X4 and MultiPlex32X8 - are pre-loaded with validated ELISA assays, available for more than 200 analytes. Different pre-loaded cartridge types provide different sample numbers, analyte numbers and internal replicates. Please refer to Object[Report,Literature,"Capillary ELISA Simple Plex Assay Menu"] and "https://www.proteinsimple.com/simple_plex_assays.html" for the list of available analytes. For the current list of available capillary ELISA cartridges, please perform Search[Model[Container,Plate,Irregular,CapillaryELISA]] with the desired CartridgeType, AnalyteMolecules or other conditions.
Default Calculation: Cartridge option is automatically set to an in-stock pre-loaded cartridge object that matches the analytes presented in the samples. If no in-stock cartridge is available for the specified options and the total number of samples is below 48, Cartridge option is automatically resolved to the customizable capillary ELISA cartridge to eliminate material waiting time. Otherwise, a pre-loaded capillary ELISA cartridge model is selected or created, with an estimated lead time of 14 days after protocol confirmation.
Pattern Description: An object of type or subtype Model[Container, Plate, Irregular, CapillaryELISA] or Object[Container, Plate, Irregular, CapillaryELISA] or Null.
Programmatic Pattern: (ObjectP[{Model[Container, Plate, Irregular, CapillaryELISA], Object[Container, Plate, Irregular, CapillaryELISA]}] | Automatic) | Null
Species
The organism (human, mouse or rat) that the samples (containing analytes of interest) are derived from. This option is used to determine the available analytes for pre-loaded capillary ELISA cartridges.
Analytes
The target(s) (e.g., peptides, proteins, antibodies, hormones) detected and quantified in each experiment sample. For customizable cartridge, only 1 analyte can be detected per sample. A pre-loaded cartridge is loaded with complete assay materials (capture antibody, detection antibody and fluorescent labelling reagent) for the selected analytes and all the samples are subject to the same analytes. For a complete list of available analytes of pre-loaded cartridges, please refer to Object[Report,Literature,"Simple Plex Assay Menu"]. To create a capillary ELISA cartridge with combination of multiple targets, please refer to Object[ManufacturingSpecification,CapillaryELISACartridge] of different analytes for more details about compatibility rules and ordering information. For SinglePlex72X1 cartridge type, only 1 analyte can be selected. For MultiAnalyte32X4 and MultiAnalyte16X4 cartridge types, up to 4 distinct analytes can be selected. For MultiPlex32X8 cartridge type, up to 8 analytes can be selected in Pro-Inflammation and Oncology panel only.
Default Calculation: If a customizable cartridge is used, Analytes option is automatically set based on the presented analyte in the Analytes field or Composition field of each input sample. If Cartridge option is set to an existing model or object of pre-loaded capillary ELISA cartridge plate, Analytes option is automatically set to the Analytes of Cartridge option. If Cartridge option is set to Null, a new pre-loaded model of capillary ELISA cartridge is requested. In this case, Analytes option is set based on the selected CartridgeType, Species and presented analytes of the input sample objects (in Analytes field or Composition field).
Programmatic Pattern: (((ObjectP[{Model[Molecule]}] | CapillaryELISAAnalyteP) | ({ObjectP[{Model[Molecule]}]..} | {CapillaryELISAAnalyteP..})) | Automatic) | Null
NumberOfReplicates
Number of times each of the input samples should be analyzed using identical experimental parameters.
Sample Preparation
SampleVolume
The initial sample amount aliquoted from the source of the input sample. If a SpikeSample is specified, this it the volume of the input sample to mix with SpikeVolume of SpikeSample before further dilution is performed.
Default Calculation: SampleVolume is automatically set to 30 Microliter when SpikeSample is not Null.
SpikeSample
For each experiment sample, the sample with known concentration(s) of analyte(s) mixed with the input sample to increase the analytical response of the unknown sample, for evaluation of sample matrices in a new ELISA assay.
SpikeVolume
The volume of the SpikeSample mixed with the SampleVolume of the input sample before further dilution is performed.
DilutionCurve
The collection of dilutions performed on each input sample (plus SpikeSample, if applicable). Dilution is required on all the loading samples for the best performance of microfluidics and the ELISA experiment is performed on all the diluted samples. For FixedDilutionVolume dilutions, the SampleAmount is the volume of the sample that is mixed with the DiluentVolume of the Diluent. For FixedDilutionFactor dilutions, the AssayVolume is the volume of the sample that is created after being diluted by the specified DilutionFactor. For a pre-loaded capillary ELISA cartridge, please refer to Object[ManufacturingSpecification,CapillaryELISACartridge] of the analytes for their recommended minimum dilution factors. For a customizable cartridge, a minimum dilution factor of 0.5 is suggested.
Figure 3.2: Use the DilutionCurve and StandardDilutionCurve option to create a collection of dilutions that will be performed on the input samples or the standard samples for capillary ELISA cartridge loading.
Default Calculation: If SerialDilutionCurve is not uploaded, DilutionCurve option is automatically set to 1 dilution with DilutionFactor of the minimum required dilution factor to generate a AssayVolume of 60 Microliter.
Programmatic Pattern: (({{GreaterEqualP[0*Microliter], GreaterEqualP[0*Microliter]}..} | {{GreaterEqualP[30*Microliter], RangeP[0, 1]}..}) | Automatic) | Null
SerialDilutionCurve
The collection of stepwise dilutions performed on each input sample (plus SpikeSample, if applicable). Dilution is required on all the loading samples for the best performance of microfluidics and the ELISA assay is performed on all the diluted samples. To use non-diluted input sample, the first dilution can be set to a dilution factor of 1. For SerialDilutionVolumes, the TransferVolume is taken out of the sample and added to a second well with the DiluentVolume of the Diluent. After thorough mixing, the TransferVolume is taken out of that well and added to a third well. This is repeated to make NumberofDilutions diluted samples. For SerialDilutionFactors, the sample is diluted by the dilution factor at each transfer step. For a pre-loaded capillary ELISA cartridge, please refer to Object[ManufacturingSpecification,CapillaryELISACartridge] of the analytes for their recommended minimum dilution factors. For a customizable cartridge, a minimum dilution factor of 0.5 is suggested.
Figure 3.3: Use the SerialDilutionCurve and StandardSerialDilutionCurve option to create a collection of stepwise dilutions that will be performed on the input samples or the standard samples for capillary ELISA cartridge loading.
Default Calculation: If DilutionCurve option is Null, SerialDilutionCurve option is automatically set to 1 NumberOfDilutions with ConstantDilutionFactor of the recommended minimum dilution factor to generate a AssayVolume of 60 Microliter.
Programmatic Pattern: (({GreaterP[0*Microliter], GreaterEqualP[30*Microliter], GreaterP[1, 1]} | {GreaterEqualP[30*Microliter], {RangeP[0, 1], GreaterP[1, 1]} | {RangeP[0, 1]..}}) | Automatic) | Null
Diluent
For each experiment sample, the diluent buffer used to mix with the input sample (plus SpikeSample, if applicable) to reduce its concentration in order to make DilutionCurve or SerialDilutionCurve. Dilution with the recommended diluent is required on all the loading samples for the best performance of microfluidics.
Default Calculation: If a pre-loaded cartridge is used for the experiment, Diluent option is automatically set to the recommended diluent provided by the assay developer for the specified Analytes or Cartridge. For a customizable cartridge, Diluent option is set to Model[Sample,"Simple Plex Sample Diluent 13"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample.
DilutionMixVolume
The volume that is pipetted up and down in the diluted input sample to mix each sample thoroughly with Diluent to make DilutionCurve or SerialDilutionCurve.
Default Calculation: DilutionMixVolume option is automatically set to half of the minimum prepared volume of the diluted sample
DilutionNumberOfMixes
The number of pipette up and down cycles that is used to mix each input sample thoroughly with Diluent to make DilutionCurve or SerialDilutionCurve.
DilutionMixRate
The speed at which the DilutionMixVolume is pipetted up and down in the diluted input sample to mix each sample thoroughly with Diluent to make DilutionCurve or SerialDilutionCurve.
Pattern Description: Greater than or equal to 0.4 microliters per second and less than or equal to 250 microliters per second.
Post Experiment
SpikeSampleStorageCondition
For each SpikeSample, the non-default conditions under which it should be stored after the protocol is completed. If left unset, the SpikeSample will be stored according to their current StorageCondition.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting, Oven} or Disposal or Null.
CaptureAntibodyStorageCondition
Indicates the condition under which the unused portion of liquid state CustomCaptureAntibody sample inside its original container is stored after the protocol is completed.
Default Calculation: CaptureAntibodyStorageCondition option is automatically set to Freezer if CaptureAntibodyResuspension is True and a customizable cartridge is used.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting, Oven} or Disposal or Null.
DetectionAntibodyStorageCondition
Indicates the condition under which the unused portion of liquid state CustomDetectionAntibody sample inside its original container is stored after the protocol is completed.
Default Calculation: DetectionAntibodyStorageCondition option is automatically set to Freezer if DetectionAntibodyResuspension is True and a customizable cartridge is used.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting, Oven} or Disposal or Null.
StandardStorageCondition
Indicates the condition under which the unused portion of liquid state Standard inside its original container is stored after the protocol is completed.
Default Calculation: StandardStorageCondition option is automatically set to Freezer if StandardResuspension is True.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting, Oven} or Disposal or Null.
StandardCaptureAntibodyStorageCondition
Indicates the condition under which the unused portion of liquid state StandardCaptureAntibody sample inside its original container is stored after the protocol is completed.
Default Calculation: StandardCaptureAntibodyStorageCondition option is automatically set to Freezer if StandardCaptureAntibodyResuspension is True and a customizable cartridge is used.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting, Oven} or Disposal or Null.
StandardDetectionAntibodyStorageCondition
Indicates the condition under which the unused portion of liquid state StandardDetectionAntibody sample inside its original container is stored after the protocol is completed.
Default Calculation: StandardDetectionAntibodyStorageCondition option is automatically set to Freezer if StandardDetectionAntibodyResuspension is True and a customizable cartridge is used.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting, Oven} or Disposal or Null.
SamplesInStorageCondition
The non-default conditions under which the SamplesIn of this experiment should be stored after the protocol is completed. If left unset, SamplesIn will be stored according to their current StorageCondition.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting, Oven} or Disposal or Null.
Capture Antibody Preparation
CustomCaptureAntibody
For each experiment sample, the capture antibody sample used in the sandwich ELISA experiment to bind with the present analytes through specific antigen-antibody interaction. The capture antibody must be labeled with digoxigenin, purified and diluted to preferably 3.5 Microgram/Milliliter before loading into the capillary ELISA cartridge. During the experiment, digoxigenin-labeled capture antibody sample first flows in the fluidic circuit to allow its binding to the immobilized anti-digoxigenin monoclonal antibody in the capillaries. Then the diluted input samples are applied to allow the binding to capture antibody for immobilization. The provided CustomCaptureAntibody can be resuspended into solution, bioconjugated with digoxigenin and diluted as desired using the related options. Please note that if the same capture antibody preparation options are specified for multiple samples (standard samples and/or input samples), the preparation of digoxigenin-labeled capture antibody is performed together. Please consider using ExperimentSamplePreparation for more customized capture antibody preparation.
Default Calculation: If a customizable cartridge is used, CustomCaptureAntibody is automatically set based on the Analytes option. The Antibodies or SecondaryAntibodies field in Model[Molecule] of Analytes is inspected. If available, the DefaultSampleModel of a digoxigenin-labeled antibody is automatically selected as CustomCaptureAntibody. If no digoxigenin-labeled antibody is available, the DefaultSampleModel of an antibody with different epitopes with CustomDetectionAntibody (if uploaded) is automatically selected as CustomCaptureAntibody. If neither CustomCaptureAntibody or CustomDetectionAntibody has been specified and two antibodies with different epitopes are available for the analyte, the DefaultSampleModel of a polyclonal antibody is preferably selected for CustomCaptureAntibody.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
CaptureAntibodyResuspension
Indicates if the CustomCaptureAntibody should be reconstituted with CaptureAntibodyResuspensionDiluent. If set to True, the original container of the CustomCaptureAntibody sample is centrifuged at 3000 RPM for 1 Minute to eliminate sample loss upon cap opening. Then the CaptureAntibodyResuspensionDiluent is delivered into the CustomCaptureAntibody's container and thorough mixing is achieved by inversion for 30 times.
Default Calculation: CaptureAntibodyResuspension is automatically set to True for a solid state CustomCaptureAntibody sample in a Customizable capillary ELISA cartridge.
CaptureAntibodyResuspensionConcentration
Default Calculation: CaptureAntibodyResuspensionConcentration option is automatically set to 1 Milligram/Milliliter if CaptureAntibodyResuspension is True and a customizable cartridge is used.
CaptureAntibodyResuspensionDiluent
The resuspension buffer mixed with the solid state CustomCaptureAntibody sample to dissolve the sample into solution.
Default Calculation: CaptureAntibodyResuspensionDiluent option is automatically set to Model[Sample, StockSolution, "Filtered PBS, Sterile"] if CaptureAntibodyResuspension is True and a customizable cartridge is used. The amount of the solid state CustomCaptureAntibody sample is used to calculate the amount of CaptureAntibodyResuspensionDiluent needed.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
CaptureAntibodyConjugation
Indicates if bioconjugation reaction between digoxigenin NHS-ester and primary amines of the CustomCaptureAntibody is required to be performed to prepare digoxigenin-labeled capture antibody sample for capillary ELISA experiment. If set to True, the conjugation mixture is vortexed at 1900 RPM for 1 Minute to allow thorough mixing and then incubated on a heat block at CaptureAntibodyConjugationTemperature for CaptureAntibodyConjugationTime to allow complete reaction.
Default Calculation: CaptureAntibodyConjugation is automatically set to True if the antibody composition of the CustomCaptureAntibody in the Customizable capillary ELISA cartridge does not show Model[Molecule,Protein,Antibody,"Anti-Digoxigenin Antibody"] as its SecondaryAntibodies.
CaptureAntibodyVolume
The volume of the unconjugated CustomCaptureAntibody sample used to react with DigoxigeninReagent to prepare digoxigenin-labeled capture antibody sample through bioconjugation synthesis.
Default Calculation: CaptureAntibodyVolume option is automatically set to 100 Microliter if CaptureAntibodyConjugation is True and a customizable cartridge is used.
DigoxigeninReagent
The digoxigenin NHS-ester reagent used to react with the primary amines of the unconjugated CustomCaptureAntibody sample to prepare digoxigenin-labeled capture antibody sample through bioconjugation synthesis.
Default Calculation: DigoxigeninReagent option is automatically set to Model[Sample,StockSolution,"Digoxigenin-NHS, 0.67 mg/mL in DMF"] if CaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
DigoxigeninReagentVolume
The volume of DigoxigeninReagent used to react with CaptureAntibodyVolume of the unconjugated CustomCaptureAntibody sample to prepare digoxigenin-labeled capture antibody sample through bioconjugation synthesis.
Default Calculation: When a customizable cartridge is used, DigoxigeninReagentVolume option is automatically set using the concentration of DigoxigeninReagent and the antibody concentration to achieve a 5:1 molar ratio between DigoxigeninReagent and the unconjugated CustomCaptureAntibody.
CaptureAntibodyConjugationBuffer
The buffer solution in which the reaction between the unconjugated CustomCaptureAntibody sample and DigoxigeninReagent happens to prepare digoxigenin-labeled capture antibody sample through bioconjugation synthesis.
Default Calculation: CaptureAntibodyConjugationBuffer option is automatically set to Model[Sample,StockSolution,"Sodium bicarbonate working stock 75 mg/mL"] if CaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
CaptureAntibodyConjugationBufferVolume
The volume of CaptureAntibodyConjugationBuffer used to mix with the unconjugated CustomCaptureAntibody sample and DigoxigeninReagent to provide a buffered environment for the bioconjugation reaction.
Default Calculation: When a customizable cartridge is used, CaptureAntibodyConjugationBufferVolume option is automatically set to 1/9 of total volume of CaptureAntibodyVolume and DigoxigeninReagentVolume.
CaptureAntibodyConjugationContainer
The container in which the unconjugated CustomCaptureAntibody sample, DigoxigeninReagent and CaptureAntibodyConjugationBuffer react to allow bioconjugation synthesis.
Default Calculation: CaptureAntibodyConjugationContainer option is automatically selected base on the total conjugation reaction volume if CaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Container] or Object[Container] or a prepared sample or Null.
Programmatic Pattern: ((ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic) | Null
CaptureAntibodyConjugationTime
The amount of reaction time that the CustomCaptureAntibody and DigoxigeninReagent are allowed to react before purification happens.
Default Calculation: CaptureAntibodyConjugationTime option is automatically set to 1 Hour if CaptureAntibodyConjugation is True and a customizable cartridge is used.
CaptureAntibodyConjugationTemperature
The temperature at which the reaction between the unconjugated CustomCaptureAntibody and DigoxigeninReagent is conducted.
Default Calculation: CaptureAntibodyConjugationTemperature option is automatically set to Ambient if CaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: Ambient or greater than or equal to -20 degrees Celsius and less than or equal to 150 degrees Celsius or Null.
CaptureAntibodyPurificationColumn
The desalting spin column used to adsorb the molecules that are smaller than its molecular weight cut-off (MWCO) on its resin bed to remove the free digoxigenin NHS-ester molecules in order to purify the synthesis product of the CustomCaptureAntibody bioconjugation reaction. The spin column is operated following the manufacturer's instruction (Figure 3.4 in ExperimentCapillaryELISA help file).
Figure 3.4: When conjugation reaction is performed on the CustomCaptureAntibody sample, a spin desalting column is used to adsorb the molecules that are smaller than its molecular weight cut-off (MWCO) on its resin bed to remove the free digoxigenin NHS-ester molecules in order to purify the synthesis product of the bioconjugation reaction. The desalting spin column is first centrifuged to remove the storage solution. Then the CaptureAntibodyColumnWashBuffer is used to remove any residues from the spin column storage solution and equilibrate the desalting spin column resins before antibody sample loading. Then the reaction product of the bioconjugation reaction is loaded into the desalting spin column and pushed through the resin bed by centrifugation. The purified conjugated CustomCaptureAntibody sample is collected for cartridge loading. For details about the operation of spin desalting columns, including the default wash buffer volumes and centrifuge times, please refer to Object[Report,Literature,"Zeba Spin Column Operation Instructions"].
Default Calculation: CaptureAntibodyPurificationColumn is automatically set to a 40K molecular weight cut-off Zeba desalting spin column depending on the total volume of the synthesis reaction, if CaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Container, Vessel, Filter] or Object[Container, Vessel, Filter] or Null.
Programmatic Pattern: (ObjectP[{Model[Container, Vessel, Filter], Object[Container, Vessel, Filter]}] | Automatic) | Null
CaptureAntibodyColumnWashBuffer
The buffer solution loaded into the CaptureAntibodyPurificationColumn after its storage buffer is removed and before the sample is loaded. The CaptureAntibodyColumnWashBuffer flushed through CaptureAntibodyPurificationColumn to remove any residues from the spin column storage solution. The washing process is repeated 3 times to equilibrate the resin bed before sample loading.
Default Calculation: CaptureAntibodyColumnWashBuffer option is automatically set to Model[Sample, StockSolution, "Filtered PBS, Sterile"] if CaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
CaptureAntibodyConjugationStorageCondition
Indicates the condition under which the unused portion of the synthesized and purified digoxigenin-labeled capture antibody sample is stored after the protocol is completed.
Default Calculation: CaptureAntibodyConjugationStorageCondition option is automatically set to Refrigerator if CaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting, Oven} or Disposal or Null.
CaptureAntibodyDilution
Indicates if digoxigenin-labeled capture antibody dilution is required before loading into capillary ELISA cartridge.
Default Calculation: When a Customizable capillary ELISA cartridge is used, CaptureAntibodyDilution is automatically set to True if CaptureAntibodyResuspension and/or CaptureAntibodyConjugation is True.
CaptureAntibodyTargetConcentration
The desired concentration of digoxigenin-labeled capture antibody after dilution. The initial mass concentration of digoxigenin-labeled capture antibody sample is acquired from the composition of the liquid state CustomCaptureAntibody sample or the CaptureAntibodyResuspension of the solid state CustomCaptureAntibody sample. A 50% concentration loss is factored in if conjugation is performed in the experiment.
Default Calculation: CaptureAntibodyTargetConcentration is automatically set to 3.5 Microgram/Milliliter if CaptureAntibodyDilution is True and a customizable cartridge is used.
CaptureAntibodyDiluent
The buffer solution used to mix with the concentrated digoxigenin-labeled capture antibody sample to achieve the desired CaptureAntibodyTargetConcentration for cartridge loading.
Default Calculation: CaptureAntibodyDiluent option is automatically set to Model[Sample, "Simple Plex Reagent Diluent"] if CaptureAntibodyDilution is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
Detection Antibody Preparation
CustomDetectionAntibody
The detection antibody sample used in the sandwich ELISA experiment. After immobilization of the analytes in the microfluidic channels, the detection antibodies bind to analyte epitopes distinct from the capture antibodies through antigen-antibody interaction. Then the streptavidin-conjugated fluorescent dyes can be attached through biotin-streptavidin interaction for data acquiring. The detection antibody must be modified with biotin, purified and diluted to preferably 3.5 Microgram/Milliliter before loading into the capillary ELISA cartridge. The provided CustomDetectionAntibody can be resuspended into solution, bioconjugated with biotin and diluted as desired using the related options. Please note that if the same detection antibody preparation options are specified for multiple samples (standard samples and/or input samples), the preparation of biotinylated detection antibody is performed together. Please consider using ExperimentSamplePreparation for more customized detection antibody preparation.
Default Calculation: If a customizable cartridge is used, CustomDetectionAntibody is automatically set based on the Analytes option. The Antibodies or SecondaryAntibodies field in Model[Molecule] of Analytes is inspected. If available, the DefaultSampleModel of a biotinylated antibody is automatically selected as CustomDetectionAntibody. If no biotinylated antibody is available, the DefaultSampleModel of an antibody with different epitopes with CustomCaptureAntibody (if specified) is automatically selected as CustomDetectionAntibody. If neither CustomCaptureAntibody or CustomDetectionAntibody has been specified and two antibodies with different epitopes are available for the analyte, the DefaultSampleModel of a monoclonal antibody is preferably selected for CustomDetectionAntibody.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
DetectionAntibodyResuspension
Indicates if the CustomDetectionAntibody should be reconstituted with DetectionAntibodyResuspensionDiluent. If set to True, the original container of the CustomDetectionAntibody sample is centrifuged at 3000 RPM for 1 Minute to eliminate sample loss upon cap opening. Then the DetectionAntibodyResuspensionDiluent is delivered into the CustomDetectionAntibody's container and thorough mixing is achieved by inversion for 30 times.
Default Calculation: DetectionAntibodyResuspension is automatically set to True for a solid state CustomDetectionAntibody sample in a Customizable capillary ELISA cartridge.
DetectionAntibodyResuspensionConcentration
Default Calculation: DetectionAntibodyResuspensionConcentration option is automatically set to 1 Milligram/Milliliter if DetectionAntibodyResuspension is True and a customizable cartridge is used.
DetectionAntibodyResuspensionDiluent
The resuspension buffer mixed with the solid state CustomDetectionAntibody sample to dissolve the sample into solution.
Default Calculation: DetectionAntibodyResuspensionDiluent option is automatically set to Model[Sample, StockSolution, "Filtered PBS, Sterile"] if DetectionAntibodyResuspension is True and a customizable cartridge is used. The amount of the solid state CustomDetectionAntibody sample is used to calculate the amount of DetectionAntibodyResuspensionDiluent needed.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
DetectionAntibodyConjugation
Indicates if bioconjugation reaction between Biotin-XX-SE (6-((6-((Biotinoyl)Amino)Hexanoyl)Amino)Hexanoic Acid, Succinimidyl Ester) and primary amines of the CustomDetectionAntibody is required to be performed to prepare biotinylated detection antibody sample for capillary ELISA experiment. If set to True, the conjugation mixture is vortexed at 1900 RPM for 1 Minute to allow thorough mixing and then incubated on a heat block at DetectionAntibodyConjugationTemperature for DetectionAntibodyConjugationTime to allow complete reaction.
Default Calculation: DetectionAntibodyConjugation is automatically set to True if the antibody composition of the CustomDetectionAntibody in the Customizable capillary ELISA cartridge does not show Model[Molecule,Protein,"Streptavidin"] as its Targets.
DetectionAntibodyVolume
The volume of the unconjugated CustomDetectionAntibody sample used to react with BiotinReagent to prepare biotinylated detection antibody sample through bioconjugation synthesis.
Default Calculation: DetectionAntibodyVolume option is automatically set to 100 Microliter if DetectionAntibodyConjugation is True and a customizable cartridge is used.
BiotinReagent
The Biotin-XX-SE (6-((6-((Biotinoyl)Amino)Hexanoyl)Amino)Hexanoic Acid, Succinimidyl Ester) or other biotinylation reagent used to mix with the unconjugated CustomDetectionAntibody sample to prepare biotinylated detection antibody sample through bioconjugation synthesis.
Default Calculation: BiotinReagent option is automatically set to Model[Sample,StockSolution,"Biotin-XX, 1 mg/mL in DMSO"] if DetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BiotinReagentVolume
The volume of BiotinReagent used to react with DetectionAntibodyVolume of the unconjugated CustomDetectionAntibody sample to prepare biotinylated detection antibody sample through bioconjugation synthesis.
Default Calculation: When a customizable cartridge is used, BiotinReagentVolume option is automatically set using the concentration of BiotinReagent and the antibody concentration to achieve a 10:1 molar ratio between BiotinReagent and the unconjugated CustomDetectionAntibody.
DetectionAntibodyConjugationBuffer
The buffer solution in which the reaction between the unconjugated CustomDetectionAntibody sample and BiotinReagent happens to prepare biotinylated detection antibody sample through bioconjugation synthesis.
Default Calculation: DetectionAntibodyConjugationBuffer option is automatically set to Model[Sample,StockSolution,"Sodium bicarbonate working stock 75 mg/mL"] if DetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
DetectionAntibodyConjugationBufferVolume
The volume of DetectionAntibodyConjugationBuffer used to mix with the unconjugated CustomDetectionAntibody sample and BiotinReagent to provide a buffered environment for the bioconjugation reaction.
Default Calculation: When a customizable cartridge is used, DetectionAntibodyConjugationBufferVolume option is automatically set to 1/9 of total volume of DetectionAntibodyVolume and BiotinReagentVolume.
DetectionAntibodyConjugationContainer
The container in which the unconjugated CustomDetectionAntibody sample, BiotinReagent and DetectionAntibodyConjugationBuffer react to allow bioconjugation synthesis.
Default Calculation: DetectionAntibodyConjugationContainer option is automatically selected base on the total conjugation reaction volume if DetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Container] or Object[Container] or a prepared sample or Null.
Programmatic Pattern: ((ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic) | Null
DetectionAntibodyConjugationTime
The amount of reaction time that the CustomDetectionAntibody and BiotinReagent are allowed to react before purification happens.
Default Calculation: DetectionAntibodyConjugationTime option is automatically set to 1 Hour if DetectionAntibodyConjugation is True and a customizable cartridge is used.
DetectionAntibodyConjugationTemperature
The temperature at which the reaction between the unconjugated CustomDetectionAntibody and BiotinReagent is conducted.
Default Calculation: DetectionAntibodyConjugationTemperature option is automatically set to Ambient if DetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: Ambient or greater than or equal to -20 degrees Celsius and less than or equal to 150 degrees Celsius or Null.
DetectionAntibodyPurificationColumn
The desalting spin column used to adsorb the molecules that are smaller than its molecular weight cut-off (MWCO) on its resin bed to remove the free biotinylation reagent molecules in order to purify the synthesis product of the CustomDetectionAntibody bioconjugation reaction. The spin column is operated following the manufacturer's instruction (Figure 3.5 in ExperimentCapillaryELISA help file).
Figure 3.5: When conjugation reaction is performed on the CustomDetectionAntibody sample, a spin desalting column is used to adsorb the molecules that are smaller than its molecular weight cut-off (MWCO) on its resin bed to remove the free digoxigenin NHS-ester molecules in order to purify the synthesis product of the bioconjugation reaction. The desalting spin column is first centrifuged to remove the storage solution. Then the DetectionAntibodyColumnWashBuffer is used to remove any residues from the spin column storage solution and equilibrate the desalting spin column resins before antibody sample loading. Then the reaction product of the bioconjugation reaction is loaded into the desalting spin column and pushed through the resin bed by centrifugation. The purified conjugated CustomDetectionAntibody sample is collected for cartridge loading. For details about the operation of spin desalting columns, including the default wash buffer volumes and centrifuge times, please refer to Object[Report,Literature,"Zeba Spin Column Operation Instructions"].
Default Calculation: DetectionAntibodyPurificationColumn is automatically set to a 40K molecular weight cut-off Zeba desalting spin column depending on the total volume of the synthesis reaction, if DetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Container, Vessel, Filter] or Object[Container, Vessel, Filter] or Null.
Programmatic Pattern: (ObjectP[{Model[Container, Vessel, Filter], Object[Container, Vessel, Filter]}] | Automatic) | Null
DetectionAntibodyColumnWashBuffer
The buffer solution loaded into the DetectionAntibodyPurificationColumn after its storage buffer is removed and before the sample is loaded. The DetectionAntibodyColumnWashBuffer flushed through DetectionAntibodyPurificationColumn to remove any residues from the spin column storage solution. The washing process is repeated 3 times to equilibrate the resin bed before sample loading.
Default Calculation: DetectionAntibodyColumnWashBuffer option is automatically set to Model[Sample, StockSolution, "Filtered PBS, Sterile"] if DetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
DetectionAntibodyConjugationStorageCondition
Indicates the condition under which the unused portion of the synthesized and purified biotinylated detection antibody sample is stored after the protocol is completed.
Default Calculation: DetectionAntibodyConjugationStorageCondition option is automatically set to Refrigerator if DetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting, Oven} or Disposal or Null.
DetectionAntibodyDilution
Indicates if biotinylated detection antibody dilution is required before loading into capillary ELISA cartridge.
Default Calculation: When a Customizable capillary ELISA cartridge is used, DetectionAntibodyDilution is automatically set to True if DetectionAntibodyResuspension and/or DetectionAntibodyConjugation is True.
DetectionAntibodyTargetConcentration
The desired concentration of biotinylated detection antibody after dilution. The initial mass concentration of biotinylated detection antibody sample is acquired from the composition of the liquid state CustomDetectionAntibody sample or the DetectionAntibodyResuspension of the solid state CustomDetectionAntibody sample. A 50% concentration loss is factored in if conjugation is performed in the experiment.
Default Calculation: DetectionAntibodyTargetConcentration is automatically set to 3.5 Microgram/Milliliter if DetectionAntibodyDilution is True and a customizable cartridge is used.
DetectionAntibodyDiluent
The buffer solution used to mix with the concentrated biotinylated detection antibody sample to achieve the desired DetectionAntibodyTargetConcentration for cartridge loading.
Default Calculation: DetectionAntibodyDiluent option is automatically set to Model[Sample, "Simple Plex Reagent Diluent"] if DetectionAntibodyDilution is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
Standard Curve
Standard
The standard samples with known concentration(s) of the analyte(s) that are used to make a dilution series and create a standard curve in the capillary ELISA experiment.
Default Calculation: If a customizable cartridge is used, Standard option is automatically set to the DefaultSampleModel of the Analytes.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
Standard Preparation
StandardResuspension
Indicates if resuspension is required to be performed on the solid state Standard sample to turn the sample into solution. If set to True, the original container of the Standard sample is centrifuged at 3000 RPM for 1 Minute to eliminate sample loss upon cap opening. Then the StandardResuspensionDiluent is delivered into the Standard's container and thorough mixing is achieved by inversion for 30 times.
Default Calculation: StandardResuspension is automatically set to True for a solid state Standard sample.
StandardResuspensionConcentration
The target concentration of the Standard sample in the solution after resuspension. The amount of the solid state Standard is used to calculate the volume of StandardResuspensionDiluent needed.
Default Calculation: If a pre-loaded cartridge is used in this protocol and the standard sample is for one of its analytes, StandardResuspensionConcentration is automatically set to 10 times of the analyte's upper limit of quantitation (ULOQ) provided by the assay developer. This information can be found in the corresponding Object[ManufacturingSpecification,CapillaryELISACartridge] of the analyte. Otherwise, StandardResuspension is automatically set to 100 Microgram/Milliliter if StandardResuspension is True.
StandardResuspensionDiluent
The resuspension buffer mixed with the solid state Standard sample to dissolve the sample into solution.
Default Calculation: If a pre-loaded cartridge is used in this protocol and the standard sample is for one of its analytes, StandardResuspensionDiluent is automatically set to the recommended diluent for this analyte suggested by the assay developer. This information can be found in the corresponding Object[ManufacturingSpecification,CapillaryELISACartridge] of the analyte. Otherwise it is automatically set to Model[Sample,"Simple Plex Sample Diluent 13"] if StandardResuspension is True.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardDilutionCurve
The collection of dilutions performed on the liquid-phase Standard sample. Dilution is required on all the loading samples for the best performance of microfluidics and the ELISA experiment is performed on all the diluted samples. Data from all measurements of all dilutions from a standard sample are used for the preparation of standard curve. For FixedDilutionVolume dilutions, the SampleAmount is the volume of the standard sample that is mixed with the DiluentVolume of the StandardDiluent. For FixedDilutionFactor dilutions, the AssayVolume is the volume of the sample that is created after being diluted by the specified DilutionFactor. For a pre-loaded capillary ELISA cartridge, please refer to Object[ManufacturingSpecification,CapillaryELISACartridge] of the analytes for their recommended minimum dilution factors. For a customizable cartridge, a minimum dilution factor of 0.5 is suggested.
Default Calculation: If Standard option is not Null and StandardSerialDilutionCurve is Null, StandardDilutionCurve is automatically set to {{60 Microliter,0.1},{60 Microliter,0.01},{60 Microliter,0.001},{60 Microliter,0.0001},{60 Microliter,0.00001}} to prepare a 60 Microliter dilution series of 5 orders.
Programmatic Pattern: (({{GreaterEqualP[0*Microliter], GreaterEqualP[0*Microliter]}..} | {{GreaterEqualP[30*Microliter], RangeP[0, 1]}..}) | Automatic) | Null
StandardSerialDilutionCurve
The collection of stepwise dilutions performed on the liquid-phase Standard sample. Dilution is required on all the loading samples for the best performance of microfluidics and the ELISA experiment is performed on all the diluted samples. Data from all measurements of all dilutions from a standard sample are used for the preparation of standard curve. To use non-diluted input sample, the first dilution can be set to a dilution factor of 1. For SerialDilutionVolumes, the TransferVolume is taken out of the sample and added to a second well with the DiluentVolume of the Diluent. After thorough mixing, the TransferVolume is taken out of that well and added to a third well. This is repeated to make NumberofDilutions diluted samples. For SerialDilutionFactors, the sample is diluted by the dilution factor at each transfer step. For a pre-loaded capillary ELISA cartridge, please refer to Object[ManufacturingSpecification,CapillaryELISACartridge] of the analytes for their recommended minimum dilution factors. For a customizable cartridge, a minimum dilution factor of 0.5 is suggested.
Default Calculation: If Standard option is uploaded and StandardDilutionCurve is uploaded, StandardSerialDilutionCurve option is automatically set to 5 NumberOfDilutions with ConstantDilutionFactor of 0.1 to generate a AssayVolume of 60 Microliter.
Programmatic Pattern: (({GreaterP[0*Microliter], GreaterEqualP[30*Microliter], GreaterP[1, 1]} | {GreaterEqualP[30*Microliter], {RangeP[0, 1], GreaterP[1, 1]} | {RangeP[0, 1]..}}) | Automatic) | Null
StandardDiluent
The diluent buffer used to mixed with the Standard sample in order to make StandardDilutionCurve or StandardSerialDilutionCurve. Dilution with the recommended diluent is required on all the loading samples for the best performance of microfluidics.
Default Calculation: If a pre-loaded cartridge is used for the experiment, StandardDiluent option is automatically set to the recommended diluent provided by the assay developer for the specified Analytes or Cartridge. For a customizable cartridge, StandardDiluent option is set to Model[Sample,"Simple Plex Sample Diluent 13"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardDilutionMixVolume
The volume that is pipetted up and down in the diluted standard sample to mix each sample thoroughly with StandardDiluent to make StandardDilutionCurve or StandardSerialDilutionCurve.
Default Calculation: StandardDilutionMixVolume is automatically set to 20 Microliter if Standard is not Null.
StandardDilutionNumberOfMixes
The number of pipette up and down cycles that is used to mix each standard sample thoroughly with StandardDiluent to make StandardDilutionCurve or StandardSerialDilutionCurve.
Default Calculation: StandardDilutionNumberOfMixes option is automatically set to 5 if Standard is not Null.
Pattern Description: Greater than or equal to 0 and less than or equal to 20 in increments of 1 or Null.
StandardDilutionMixRate
The speed at which the StandardDilutionMixVolume is pipetted up and down in the diluted standard sample to mix each sample thoroughly with StandardDiluent to make StandardDilutionCurve or StandardSerialDilutionCurve.
Default Calculation: StandardDilutionMixRate option is automatically set to 100 Microliter/Second if Standard is not Null.
Pattern Description: Greater than or equal to 0.4 microliters per second and less than or equal to 250 microliters per second or Null.
Standard Capture Antibody Preparation
StandardCaptureAntibody
The standard capture antibody sample used in the sandwich ELISA experiment to bind with analytes in the standard sample through specific antigen-antibody interaction. The standard capture antibody must be labeled with digoxigenin, purified and diluted to preferably 3.5 Microgram/Milliliter before loading into the capillary ELISA cartridge. During the experiment, digoxigenin-labeled standard capture antibody sample first flows in the fluidic circuit to allow its binding to the immobilized anti-digoxigenin monoclonal antibody in the capillaries. Then the diluted standard samples are applied to allow the binding to standard capture antibody for immobilization. The provided StandardCaptureAntibody can be resuspended into solution, bioconjugated with digoxigenin and diluted as desired using the related options. Please note that if the same capture antibody preparation options are specified for multiple samples (standard samples and/or input samples), the preparation of digoxigenin-labeled capture antibody is performed together. Please consider using ExperimentSamplePreparation for more customized standard capture antibody preparation.
Default Calculation: If a customizable cartridge is used, StandardCaptureAntibody is automatically set based on the presented analyte (from Analytes option) in the standard sample. The Antibodies or SecondaryAntibodies field in Model[Molecule] of the analyte is inspected. If available, the DefaultSampleModel of a digoxigenin-labeled antibody is automatically selected as StandardCaptureAntibody. If no digoxigenin-labeled antibody is available, the DefaultSampleModel of an antibody with different epitopes with StandardDetectionAntibody (if uploaded) is automatically selected as StandardCaptureAntibody. If neither StandardCaptureAntibody or StandardDetectionAntibody has been specified and two antibodies with different epitopes are available for the analyte, the DefaultSampleModel of a polyclonal antibody is preferably selected for StandardCaptureAntibody.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardCaptureAntibodyResuspension
Indicates if the StandardCaptureAntibody should be reconstituted with StandardCaptureAntibodyResuspensionDiluent. If set to True, the original container of the StandardCaptureAntibody sample is centrifuged at 3000 RPM for 1 Minute to eliminate sample loss upon cap opening. Then the StandardCaptureAntibodyResuspensionDiluent is delivered into the StandardCaptureAntibody's container and thorough mixing is achieved by inversion for 30 times.
Default Calculation: StandardCaptureAntibodyResuspension is automatically set to True for a solid state StandardCaptureAntibody sample in a customizable cartridge.
StandardCaptureAntibodyResuspensionConcentration
Default Calculation: StandardCaptureAntibodyResuspensionConcentration option is automatically set to 1 Milligram/Milliliter if StandardCaptureAntibodyResuspension is True and a customizable cartridge is used.
StandardCaptureAntibodyResuspensionDiluent
The resuspension buffer mixed with the solid state StandardCaptureAntibody sample to dissolve the sample into solution.
Default Calculation: StandardCaptureAntibodyResuspensionDiluent option is automatically set to Model[Sample, StockSolution, "Filtered PBS, Sterile"] if StandardCaptureAntibodyResuspension is True and a customizable cartridge is used. The amount of the solid state StandardCaptureAntibody sample is used to calculate the amount of StandardCaptureAntibodyResuspensionDiluent needed.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardCaptureAntibodyConjugation
Indicates if bioconjugation reaction between digoxigenin NHS-ester and primary amines of the StandardCaptureAntibody is required to be performed to prepare digoxigenin-labeled standard capture antibody sample for capillary ELISA experiment. If set to True, the conjugation mixture is vortexed at 1900 RPM for 1 Minute to allow thorough mixing and then incubated on a heat block at StandardCaptureAntibodyConjugationTemperature for StandardCaptureAntibodyConjugationTime to allow complete reaction.
Default Calculation: StandardCaptureAntibodyConjugation is automatically set to True if the antibody composition of the StandardCaptureAntibody in the Customizable capillary ELISA cartridge does not show Model[Molecule,Protein,Antibody,"Anti-Digoxigenin Antibody"] as its SecondaryAntibodies.
StandardCaptureAntibodyVolume
The volume of the unconjugated StandardCaptureAntibody sample used to react with StandardDigoxigeninReagent to prepare digoxigenin-labeled standard capture antibody sample through bioconjugation synthesis.
Default Calculation: StandardCaptureAntibodyVolume option is automatically set to 100 Microliter if StandardCaptureAntibodyConjugation is True and a customizable cartridge is used.
StandardDigoxigeninReagent
The digoxigenin NHS-ester reagent used to react with the primary amines of the unconjugated StandardCaptureAntibody sample to prepare digoxigenin-labeled standard capture antibody sample through bioconjugation synthesis.
Default Calculation: StandardDigoxigeninReagent option is automatically set to Model[Sample,StockSolution,"Digoxigenin-NHS, 0.67 mg/mL in DMF"] if StandardCaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardDigoxigeninReagentVolume
The volume of StandardDigoxigeninReagent used to react with StandardCaptureAntibodyVolume of the unconjugated StandardCaptureAntibody sample to prepare digoxigenin-labeled capture antibody sample through bioconjugation synthesis.
Default Calculation: When a customizable cartridge is used, StandardDigoxigeninReagentVolume option is automatically set using the concentration of StandardDigoxigeninReagent and the antibody concentration to achieve a 5:1 molar ratio between StandardDigoxigeninReagent and the unconjugated StandardCaptureAntibody.
StandardCaptureAntibodyConjugationBuffer
The buffer solution in which the reaction between the unconjugated StandardCaptureAntibody sample and StandardDigoxigeninReagent happens to prepare digoxigenin-labeled capture antibody sample through bioconjugation synthesis.
Default Calculation: StandardCaptureAntibodyConjugationBuffer option is automatically set to Model[Sample,StockSolution,"Sodium bicarbonate working stock 75 mg/mL"] if StandardCaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardCaptureAntibodyConjugationBufferVolume
The volume of StandardCaptureAntibodyConjugationBuffer used to mix with the unconjugated StandardCaptureAntibody sample and StandardDigoxigeninReagent to provide a buffered environment for the bioconjugation reaction.
Default Calculation: When a customizable cartridge is used, StandardCaptureAntibodyConjugationBufferVolume option is automatically set to 1/9 of total volume of StandardCaptureAntibodyVolume and StandardDigoxigeninReagentVolume.
StandardCaptureAntibodyConjugationContainer
The container in which the unconjugated StandardCaptureAntibody sample, StandardDigoxigeninReagent and StandardCaptureAntibodyConjugationBuffer react to allow bioconjugation synthesis.
Default Calculation: StandardCaptureAntibodyConjugationContainer option is automatically selected base on the total conjugation reaction volume if StandardCaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Container] or Object[Container] or a prepared sample or Null.
Programmatic Pattern: ((ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic) | Null
StandardCaptureAntibodyConjugationTime
The amount of reaction time that the StandardCaptureAntibody and StandardDigoxigeninReagent are allowed to react before purification happens.
Default Calculation: StandardCaptureAntibodyConjugationTime option is automatically set to 1 Hour if StandardCaptureAntibodyConjugation is True and a customizable cartridge is used.
StandardCaptureAntibodyConjugationTemperature
The temperature at which the reaction between the unconjugated StandardCaptureAntibody and StandardDigoxigeninReagent is conducted.
Default Calculation: StandardCaptureAntibodyConjugationTemperature option is automatically set to Ambient if StandardCaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: Ambient or greater than or equal to -20 degrees Celsius and less than or equal to 150 degrees Celsius or Null.
StandardCaptureAntibodyPurificationColumn
The desalting spin column used to adsorb the molecules that are smaller than its molecular weight cut-off (MWCO) on its resin bed to remove the free digoxigenin NHS-ester molecules in order to purify the synthesis product of the StandardCaptureAntibody bioconjugation reaction. The spin column is operated following the manufacturer's instruction (Figure 3.6 in ExperimentCapillaryELISA help file).
Figure 3.6: When conjugation reaction is performed on the StandardCaptureAntibody sample, a spin desalting column is used to adsorb the molecules that are smaller than its molecular weight cut-off (MWCO) on its resin bed to remove the free digoxigenin NHS-ester molecules in order to purify the synthesis product of the bioconjugation reaction. The desalting spin column is first centrifuged to remove the storage solution. Then the StandardCaptureAntibodyColumnWashBuffer is used to remove any residues from the spin column storage solution and equilibrate the desalting spin column resins before antibody sample loading. Then the reaction product of the bioconjugation reaction is loaded into the desalting spin column and pushed through the resin bed by centrifugation. The purified conjugated StandardCaptureAntibody sample is collected for cartridge loading. For details about the operation of spin desalting columns, including the default wash buffer volumes and centrifuge times, please refer to Object[Report,Literature,"Zeba Spin Column Operation Instructions"].
Default Calculation: StandardCaptureAntibodyPurificationColumn is automatically set to a 40K molecular weight cut-off Zeba desalting spin column depending on the total volume of the synthesis reaction, if StandardCaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Container, Vessel, Filter] or Object[Container, Vessel, Filter] or Null.
Programmatic Pattern: (ObjectP[{Model[Container, Vessel, Filter], Object[Container, Vessel, Filter]}] | Automatic) | Null
StandardCaptureAntibodyColumnWashBuffer
The buffer solution loaded into the StandardCaptureAntibodyPurificationColumn after its storage buffer is removed and before the sample is loaded. The StandardCaptureAntibodyColumnWashBuffer flushed through StandardCaptureAntibodyPurificationColumn to remove any residues from the spin column storage solution. The washing process is repeated 3 times to equilibrate the resin bed before sample loading.
Default Calculation: StandardCaptureAntibodyColumnWashBuffer option is automatically set to Model[Sample, StockSolution, "Filtered PBS, Sterile"] if StandardCaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardCaptureAntibodyConjugationStorageCondition
Indicates the condition under which the unused portion of the synthesized and purified digoxigenin-labeled standard capture antibody sample is stored after the protocol is completed.
Default Calculation: StandardCaptureAntibodyConjugationStorageCondition option is automatically set to Refrigerator if StandardCaptureAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting, Oven} or Disposal or Null.
StandardCaptureAntibodyDilution
Indicates if digoxigenin-labeled standard capture antibody dilution is required before loading into capillary ELISA cartridge.
Default Calculation: When a Customizable capillary ELISA cartridge is used, StandardCaptureAntibodyDilution is automatically set to True if StandardCaptureAntibodyResuspension and/or StandardCaptureAntibodyConjugation is True.
StandardCaptureAntibodyTargetConcentration
The desired concentration of digoxigenin-labeled standard capture antibody after dilution. The initial mass concentration of digoxigenin-labeled standard capture antibody sample is acquired from the composition of the liquid state StandardCaptureAntibody sample or the StandardCaptureAntibodyResuspension of the solid state StandardCaptureAntibody sample. A 50% concentration loss is factored in if conjugation is performed in the experiment.
Default Calculation: StandardCaptureAntibodyTargetConcentration is automatically set to 3.5 Microgram/Milliliter if StandardCaptureAntibodyDilution is True and a customizable cartridge is used.
StandardCaptureAntibodyDiluent
The buffer solution used to mix with the concentrated digoxigenin-labeled standard capture antibody sample to achieve the desired StandardCaptureAntibodyTargetConcentration for cartridge loading.
Default Calculation: StandardCaptureAntibodyDiluent option is automatically set to Model[Sample, "Simple Plex Reagent Diluent"] if StandardCaptureAntibodyDilution is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
Standard Detection Antibody Preparation
StandardDetectionAntibody
The standard detection antibody sample used in the sandwich ELISA experiment. After immobilization of the analytes in the microfluidic channels, the standard detection antibodies bind to analyte epitopes distinct from the standard capture antibodies through antigen-antibody interaction. Then the streptavidin-conjugated fluorescent dyes can be attached through biotin-streptavidin interaction for data acquiring. The standard detection antibody must be modified with biotin, purified and diluted to preferably 3.5 Microgram/Milliliter before loading into the capillary ELISA cartridge. The provided StandardDetectionAntibody can be resuspended into solution, bioconjugated with biotin and diluted as desired using the related options. Please note that if the same detection antibody preparation options are specified for multiple samples (standard samples and/or input samples), the preparation of biotinylated detection antibody is performed together. Please consider using ExperimentSamplePreparation for more customized standard detection antibody preparation.
Default Calculation: If a customizable cartridge is used, StandardDetectionAntibody is automatically set based on the presented analyte (from Analytes option) in the standard sample. The Antibodies or SecondaryAntibodies field in Model[Molecule] of the main analyte is inspected. If available, the DefaultSampleModel of a biotinylated antibody is automatically selected as StandardDetectionAntibody. If no biotinylated antibody is available, the DefaultSampleModel of an antibody with different epitopes with StandardCaptureAntibody (if specified) is automatically selected as CustomDetectionAntibody. If neither StandardCaptureAntibody or StandardDetectionAntibody has been specified and two antibodies with different epitopes are available for the analyte, the DefaultSampleModel of a monoclonal antibody is preferably selected for StandardDetectionAntibody.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardDetectionAntibodyResuspension
Indicates if the StandardCaptureAntibody should be reconstituted with StandardCaptureAntibodyResuspensionDiluent. If set to True, the original container of the StandardDetectionAntibody sample is centrifuged at 3000 RPM for 1 Minute to eliminate sample loss upon cap opening. Then the StandardDetectionAntibodyResuspensionDiluent is delivered into the StandardDetectionAntibody's container and thorough mixing is achieved by inversion for 30 times.
Default Calculation: StandardDetectionAntibodyResuspension is automatically set to True for a solid state StandardDetectionAntibody sample in a Customizable capillary ELISA cartridge.
StandardDetectionAntibodyResuspensionConcentration
Default Calculation: StandardDetectionAntibodyResuspensionConcentration option is automatically set to 1 Milligram/Milliliter if StandardDetectionAntibodyResuspension is True and a customizable cartridge is used.
StandardDetectionAntibodyResuspensionDiluent
The resuspension buffer mixed with the solid state StandardDetectionAntibody sample to dissolve the sample into solution.
Default Calculation: StandardDetectionAntibodyResuspensionDiluent option is automatically set to Model[Sample, StockSolution, "Filtered PBS, Sterile"] if StandardDetectionAntibodyResuspension is True and a customizable cartridge is used. The amount of the solid state StandardDetectionAntibody sample is used to calculate the amount of StandardDetectionAntibodyResuspensionDiluent needed.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardDetectionAntibodyConjugation
Indicates if bioconjugation reaction between Biotin-XX-SE (6-((6-((Biotinoyl)Amino)Hexanoyl)Amino)Hexanoic Acid, Succinimidyl Ester) and primary amines of the StandardDetectionAntibody is required to be performed to prepare biotinylated detection antibody sample for capillary ELISA experiment. If set to True, the conjugation mixture is vortexed at 1900 RPM for 1 Minute to allow thorough mixing and then incubated on a heat block at StandardDetectionAntibodyConjugationTemperature for StandardDetectionAntibodyConjugationTime to allow complete reaction.
Default Calculation: StandardDetectionAntibodyConjugation is automatically set to True if the antibody composition of the StandardDetectionAntibody in the Customizable capillary ELISA cartridge does not show Model[Molecule,Protein,"Streptavidin"] as its Targets.
StandardDetectionAntibodyVolume
The volume of the unconjugated standard detection antibody sample used to react with StandardBiotinReagent to prepare biotinylated standard detection antibody sample through bioconjugation synthesis.
Default Calculation: StandardDetectionAntibodyVolume option is automatically set to 100 Microliter if StandardDetectionAntibodyConjugation is True and a customizable cartridge is used.
StandardBiotinReagent
The Biotin-XX-SE (6-((6-((Biotinoyl)Amino)Hexanoyl)Amino)Hexanoic Acid, Succinimidyl Ester) or other biotinylation reagent used to mix with the unconjugated StandardDetectionAntibody sample to prepare biotinylated standard detection antibody sample through bioconjugation synthesis.
Default Calculation: StandardBiotinReagent option is automatically set to Model[Sample,StockSolution,"Biotin-XX, 1 mg/mL in DMSO"] if StandardDetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardBiotinReagentVolume
The volume of StandardBiotinReagent used to react with StandardDetectionAntibodyVolume of the unconjugated StandardDetectionAntibody sample to prepare biotinylated detection antibody sample through bioconjugation synthesis.
Default Calculation: When a customizable cartridge is used, StandardBiotinReagentVolume option is automatically set using the concentration of StandardBiotinReagent and the antibody concentration to achieve a 10:1 molar ratio between StandardBiotinReagent and the unconjugated StandardDetectionAntibody.
StandardDetectionAntibodyConjugationBuffer
The buffer solution in which the reaction between the unconjugated StandardDetectionAntibody sample and StandardBiotinReagent happens to prepare biotinylated standard detection antibody sample through bioconjugation synthesis.
Default Calculation: StandardDetectionAntibodyConjugationBuffer option is automatically set to Model[Sample,StockSolution,"Sodium bicarbonate working stock 75 mg/mL"] if StandardDetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardDetectionAntibodyConjugationBufferVolume
The volume of StandardDetectionAntibodyConjugationBuffer used to mix with the unconjugated StandardDetectionAntibody sample and StandardBiotinReagent to provide a buffered environment for the bioconjugation reaction.
Default Calculation: When a customizable cartridge is used, StandardDetectionAntibodyConjugationBufferVolume option is automatically set to 1/9 of total volume of StandardDetectionAntibodyVolume and StandardBiotinReagentVolume if both are uploaded.
StandardDetectionAntibodyConjugationContainer
The container in which the unconjugated StandardDetectionAntibody sample, StandardBiotinReagent and StandardDetectionAntibodyConjugationBuffer react to allow bioconjugation synthesis.
Default Calculation: StandardDetectionAntibodyConjugationContainer option is automatically selected base on the total conjugation reaction volume if StandardDetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Container] or Object[Container] or a prepared sample or Null.
Programmatic Pattern: ((ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic) | Null
StandardDetectionAntibodyConjugationTime
The amount of reaction time that the StandardDetectionAntibody and StandardBiotinReagent are allowed to react before purification happens.
Default Calculation: StandardDetectionAntibodyConjugationTime option is automatically set to 1 Hour if StandardDetectionAntibodyConjugation is True and a customizable cartridge is used.
StandardDetectionAntibodyConjugationTemperature
The temperature at which the reaction between the unconjugated StandardDetectionAntibody and StandardBiotinReagent is conducted.
Default Calculation: StandardDetectionAntibodyConjugationTemperature option is automatically set to Ambient if StandardDetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: Ambient or greater than or equal to -20 degrees Celsius and less than or equal to 150 degrees Celsius or Null.
StandardDetectionAntibodyPurificationColumn
The desalting spin column used to adsorb the molecules that are smaller than its molecular weight cut-off (MWCO) on its resin bed to remove the free biotinylation reagent molecules in order to purify the synthesis product of the StandardDetectionAntibody bioconjugation reaction. The spin column is operated following the manufacturer's instruction (Figure 3.7 in ExperimentCapillaryELISA help file).
Figure 3.7: When conjugation reaction is performed on the StandardDetectionAntibody sample, a spin desalting column is used to adsorb the molecules that are smaller than its molecular weight cut-off (MWCO) on its resin bed to remove the free digoxigenin NHS-ester molecules in order to purify the synthesis product of the bioconjugation reaction. The desalting spin column is first centrifuged to remove the storage solution. Then the StandardDetectionAntibodyColumnWashBuffer is used to remove any residues from the spin column storage solution and equilibrate the desalting spin column resins before antibody sample loading. Then the reaction product of the bioconjugation reaction is loaded into the desalting spin column and pushed through the resin bed by centrifugation. The purified conjugated StandardDetectionAntibody sample is collected for cartridge loading. For details about the operation of spin desalting columns, including the default wash buffer volumes and centrifuge times, please refer to Object[Report,Literature,"Zeba Spin Column Operation Instructions"].
Default Calculation: StandardDetectionAntibodyPurificationColumn is automatically set to a 40K molecular weight cut-off Zeba desalting spin column depending on the total volume of the synthesis reaction, if StandardDetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Container, Vessel, Filter] or Object[Container, Vessel, Filter] or Null.
Programmatic Pattern: (ObjectP[{Model[Container, Vessel, Filter], Object[Container, Vessel, Filter]}] | Automatic) | Null
StandardDetectionAntibodyColumnWashBuffer
The buffer solution loaded into the StandardDetectionAntibodyPurificationColumn after its storage buffer is removed and before the sample is loaded. The StandardDetectionAntibodyColumnWashBuffer flushed through DetectionAntibodyPurificationColumn to remove any residues from the spin column storage solution. The washing process is repeated 3 times to equilibrate the resin bed before sample loading.
Default Calculation: StandardDetectionAntibodyColumnWashBuffer option is automatically set to Model[Sample, StockSolution, "Filtered PBS, Sterile"] if StandardDetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardDetectionAntibodyConjugationStorageCondition
Indicates the condition under which the unused portion of the synthesized and purified biotinylated detection antibody sample is stored after the protocol is completed.
Default Calculation: StandardDetectionAntibodyConjugationStorageCondition option is automatically set to Refrigerator if StandardDetectionAntibodyConjugation is True and a customizable cartridge is used.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting, Oven} or Disposal or Null.
StandardDetectionAntibodyDilution
Indicates if biotinylated standard detection antibody dilution is required before loading into capillary ELISA cartridge.
Default Calculation: When a Customizable capillary ELISA cartridge is used, StandardDetectionAntibodyDilution is automatically set to True if StandardDetectionAntibodyResuspension and/or StandardDetectionAntibodyConjugation is True.
StandardDetectionAntibodyTargetConcentration
The desired concentration of biotinylated standard detection antibody after dilution. The initial mass concentration of biotinylated standard detection antibody sample is acquired from the composition of the liquid state StandardDetectionAntibody sample or the StandardDetectionAntibodyResuspension of the solid state StandardDetectionAntibody sample. A 50% concentration loss is factored in if conjugation is performed in the experiment.
Default Calculation: StandardDetectionAntibodyTargetConcentration is automatically set to 3.5 Microgram/Milliliter if StandardDetectionAntibodyDilution is True and a customizable cartridge is used.
StandardDetectionAntibodyDiluent
The buffer solution used to mix with the concentrated biotinylated detection antibody sample to achieve the specified StandardDetectionAntibodyTargetConcentration for cartridge loading.
Default Calculation: StandardDetectionAntibodyDiluent option is automatically set to Model[Sample, "Simple Plex Reagent Diluent"] if StandardDetectionAntibodyDilution is True and a customizable cartridge is used.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
Washing
WashBuffer
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample.
Cartridge Loading
LoadingVolume
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 90 microliters.
CaptureAntibodyLoadingVolume
The volume of the digoxigenin-labeled capture antibody sample loaded into the capture antibody well of the customizable capillaryELISA cartridge plate.
Default Calculation: CaptureAntibodyLoadingVolume option is automatically set to 50 Microliter if a customizable cartridge is used.
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 50 microliters or Null.
DetectionAntibodyLoadingVolume
The volume of the biotinylated detection antibody sample to be loaded into the detection antibody well of the customizable capillaryELISA cartridge plate.
Default Calculation: DetectionAntibodyLoadingVolume option is automatically set to 50 Microliter iif a customizable cartridge is used.
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 50 microliters or Null.
StandardLoadingVolume
Default Calculation: StandardLoadingVolume option is automatically set to 50 Microliter if Standard option is not Null.
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 90 microliters or Null.
StandardCaptureAntibodyLoadingVolume
The volume of the digoxigenin-labeled standard capture antibody sample to be loaded into the capture antibody well of the customizable capillaryELISA cartridge plate.
Default Calculation: StandardCaptureAntibodyLoadingVolume option is automatically set to 50 Microliter if a customizable cartridge is used.
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 50 microliters or Null.
StandardDetectionAntibodyLoadingVolume
The volume of the biotinylated standard detection antibody sample to be loaded into the detection antibody well of the customizable capillaryELISA cartridge plate.
Default Calculation: StandardDetectionAntibodyLoadingVolume option is automatically set to 50 Microliter if a customizable cartridge is used.
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 50 microliters or Null.
Data Processing
StandardComposition
The known concentration(s) of presented analyte(s) in the Standard sample before dilution. This information can be used to generate standard curve.
Default Calculation: KnownConcentration is automatically set to StandardResuspensionConcentration if the resuspension is performed within this protocol. Otherwise it is automatically set using the information from the Composition field of the Standard sample.
Programmatic Pattern: ({{GreaterP[0*Molar] | GreaterP[0*(Gram/Liter)], ObjectP[{Model[Molecule]}]}..} | Automatic) | Null
SpikeConcentration
The known concentration(s) of the analyte(s) in SpikeSample. This information is used for calculation of the analyte concentration(s) in unknown sample if a standard curve is available. For a pre-loaded cartridge with more than 1 analyte, concentrations of different analytes can be provided for the SpikeSample.
Default Calculation: SpikeConcentration is automatically set using the information from the Composition field of the SpikeSample.
Programmatic Pattern: ({{GreaterP[0*Molar] | GreaterP[0*(Gram/Liter)], ObjectP[{Model[Molecule]}]}..} | Automatic) | Null
Model Input
PreparedModelContainer
Indicates the container in which a Model[Sample] specified as input to the experiment function will be prepared.
Default Calculation: If PreparedModelAmount is set to All and when the input model has a product associated with both Amount and DefaultContainerModel populated, automatically set to the DefaultContainerModel value in the product. Otherwise set to Model[Container, Vessel, "2mL Tube"].
PreparedModelAmount
Indicates the amount of a Model[Sample] specified as input to the experiment function that will be prepared in the PreparedModelContainer. When set to All and the input model sample is not preparable, the entire amount of the input model sample that comes from one of the Products is prepared. The selected product must have both Amount and DefaultContainerModel populated, and it must not be a KitProduct. When set to All and the input model is preparable such as water, 1 Milliliter of the input model sample is prepared.
Sample Prep Options
Sample Preparation
PreparatoryUnitOperations
Specifies a sequence of transferring, aliquoting, consolidating, or mixing of new or existing samples before the main experiment. These prepared samples can be used in the main experiment by referencing their defined name. For more information, please reference the documentation for ExperimentSamplePreparation.
Pattern Description: List of one or more unit Operation ManualSamplePreparation or RoboticSamplePreparation or unit Operation must match SamplePreparationP entries or Null.
Programmatic Pattern: {((ManualSamplePreparationMethodP | RoboticSamplePreparationMethodP) | SamplePreparationP)..} | Null
Preparatory Incubation
Incubate
Indicates if the SamplesIn should be incubated at a fixed temperature prior to starting the experiment or any aliquoting. Sample Preparation occurs in the order of Incubation, Centrifugation, Filtration, and then Aliquoting (if specified).
Default Calculation: Resolves to True if any of the corresponding Incubation options are set. Otherwise, resolves to False.
IncubationTemperature
Temperature at which the SamplesIn should be incubated for the duration of the IncubationTime prior to starting the experiment.
Pattern Description: Ambient or greater than or equal to -20 degrees Celsius and less than or equal to 500 degrees Celsius or Null.
Programmatic Pattern: ((Ambient | RangeP[$MinIncubationTemperature, $MaxIncubationTemperature]) | Automatic) | Null
IncubationTime
Duration for which SamplesIn should be incubated at the IncubationTemperature, prior to starting the experiment.
Mix
Default Calculation: Automatically resolves to True if any Mix related options are set. Otherwise, resolves to False.
MixType
Default Calculation: Automatically resolves based on the container of the sample and the Mix option.
Pattern Description: Roll, Vortex, Sonicate, Pipette, Invert, Stir, Shake, Homogenize, Swirl, Disrupt, or Nutate or Null.
MixUntilDissolved
Indicates if the mix should be continued up to the MaxIncubationTime or MaxNumberOfMixes (chosen according to the mix Type), in an attempt dissolve any solute. Any mixing/incubation will occur prior to starting the experiment.
Default Calculation: Automatically resolves to True if MaxIncubationTime or MaxNumberOfMixes is set.
MaxIncubationTime
Maximum duration of time for which the samples will be mixed while incubated in an attempt to dissolve any solute, if the MixUntilDissolved option is chosen. This occurs prior to starting the experiment.
Default Calculation: Automatically resolves based on MixType, MixUntilDissolved, and the container of the given sample.
IncubationInstrument
Default Calculation: Automatically resolves based on the options Mix, Temperature, MixType and container of the sample.
Pattern Description: An object of type or subtype Model[Instrument, Roller], Model[Instrument, OverheadStirrer], Model[Instrument, Vortex], Model[Instrument, Shaker], Model[Instrument, BottleRoller], Model[Instrument, Roller], Model[Instrument, Sonicator], Model[Instrument, HeatBlock], Model[Instrument, Homogenizer], Model[Instrument, Disruptor], Model[Instrument, Nutator], Model[Instrument, Thermocycler], Model[Instrument, EnvironmentalChamber], Model[Instrument, Pipette], Object[Instrument, Roller], Object[Instrument, OverheadStirrer], Object[Instrument, Vortex], Object[Instrument, Shaker], Object[Instrument, BottleRoller], Object[Instrument, Roller], Object[Instrument, Sonicator], Object[Instrument, HeatBlock], Object[Instrument, Homogenizer], Object[Instrument, Disruptor], Object[Instrument, Nutator], Object[Instrument, Thermocycler], Object[Instrument, EnvironmentalChamber], or Object[Instrument, Pipette] or Null.
AnnealingTime
Minimum duration for which the SamplesIn should remain in the incubator allowing the system to settle to room temperature after the IncubationTime has passed but prior to starting the experiment.
IncubateAliquotContainer
The desired type of container that should be used to prepare and house the incubation samples which should be used in lieu of the SamplesIn for the experiment.
Programmatic Pattern: ((ObjectP[Model[Container]] | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
IncubateAliquotDestinationWell
The desired position in the corresponding IncubateAliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
IncubateAliquot
The amount of each sample that should be transferred from the SamplesIn into the IncubateAliquotContainer when performing an aliquot before incubation.
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container.
Pattern Description: All or greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
Preparatory Centrifugation
Centrifuge
Indicates if the SamplesIn should be centrifuged prior to starting the experiment or any aliquoting. Sample Preparation occurs in the order of Incubation, Centrifugation, Filtration, and then Aliquoting (if specified).
Default Calculation: Resolves to True if any of the corresponding Centrifuge options are set. Otherwise, resolves to False.
CentrifugeInstrument
Pattern Description: An object of type or subtype Model[Instrument, Centrifuge] or Object[Instrument, Centrifuge] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, Centrifuge], Object[Instrument, Centrifuge]}] | Automatic) | Null
CentrifugeIntensity
The rotational speed or the force that will be applied to the samples by centrifugation prior to starting the experiment.
Pattern Description: Greater than 0 revolutions per minute or greater than 0 standard accelerations due to gravity on the surface of the earth or Null.
Programmatic Pattern: ((GreaterP[0*RPM] | GreaterP[0*GravitationalAcceleration]) | Automatic) | Null
CentrifugeTime
CentrifugeTemperature
The temperature at which the centrifuge chamber should be held while the samples are being centrifuged prior to starting the experiment.
Pattern Description: Ambient or greater than or equal to -10 degrees Celsius and less than or equal to 40 degrees Celsius or Null.
CentrifugeAliquotContainer
The desired type of container that should be used to prepare and house the centrifuge samples which should be used in lieu of the SamplesIn for the experiment.
Programmatic Pattern: ((ObjectP[Model[Container]] | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
CentrifugeAliquotDestinationWell
The desired position in the corresponding AliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
CentrifugeAliquot
The amount of each sample that should be transferred from the SamplesIn into the CentrifugeAliquotContainer when performing an aliquot before centrifugation.
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container.
Pattern Description: All or greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
Preparatory Filtering
Filtration
Indicates if the SamplesIn should be filter prior to starting the experiment or any aliquoting. Sample Preparation occurs in the order of Incubation, Centrifugation, Filtration, and then Aliquoting (if specified).
Default Calculation: Resolves to True if any of the corresponding Filter options are set. Otherwise, resolves to False.
FiltrationType
Default Calculation: Will automatically resolve to a filtration type appropriate for the volume of sample being filtered.
FilterInstrument
Default Calculation: Will automatically resolved to an instrument appropriate for the filtration type.
Pattern Description: An object of type or subtype Model[Instrument, FilterBlock], Object[Instrument, FilterBlock], Model[Instrument, PeristalticPump], Object[Instrument, PeristalticPump], Model[Instrument, VacuumPump], Object[Instrument, VacuumPump], Model[Instrument, Centrifuge], Object[Instrument, Centrifuge], Model[Instrument, SyringePump], or Object[Instrument, SyringePump] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, FilterBlock], Object[Instrument, FilterBlock], Model[Instrument, PeristalticPump], Object[Instrument, PeristalticPump], Model[Instrument, VacuumPump], Object[Instrument, VacuumPump], Model[Instrument, Centrifuge], Object[Instrument, Centrifuge], Model[Instrument, SyringePump], Object[Instrument, SyringePump]}] | Automatic) | Null
Filter
The filter that should be used to remove impurities from the SamplesIn prior to starting the experiment.
Default Calculation: Will automatically resolve to a filter appropriate for the filtration type and instrument.
Pattern Description: An object of type or subtype Model[Container, Plate, Filter], Model[Container, Vessel, Filter], or Model[Item, Filter] or Null.
Programmatic Pattern: (ObjectP[{Model[Container, Plate, Filter], Model[Container, Vessel, Filter], Model[Item, Filter]}] | Automatic) | Null
FilterMaterial
The membrane material of the filter that should be used to remove impurities from the SamplesIn prior to starting the experiment.
Default Calculation: Resolves to an appropriate filter material for the given sample is Filtration is set to True.
Pattern Description: Cellulose, Cotton, Polyethylene, Polypropylene, PTFE, Nylon, PES, PLUS, PVDF, GlassFiber, GHP, UHMWPE, EPDM, DuraporePVDF, GxF, ZebaDesaltingResin, NickelResin, AgaroseResin, CobaltResin, Silica, HLB, or AnoporeAlumina or Null.
PrefilterMaterial
The material from which the prefilter filtration membrane should be made of to remove impurities from the SamplesIn prior to starting the experiment.
Pattern Description: Cellulose, Cotton, Polyethylene, Polypropylene, PTFE, Nylon, PES, PLUS, PVDF, GlassFiber, GHP, UHMWPE, EPDM, DuraporePVDF, GxF, ZebaDesaltingResin, NickelResin, AgaroseResin, CobaltResin, Silica, HLB, or AnoporeAlumina or Null.
FilterPoreSize
The pore size of the filter that should be used when removing impurities from the SamplesIn prior to starting the experiment.
Default Calculation: Resolves to an appropriate filter pore size for the given sample is Filtration is set to True.
Pattern Description: 0.008 micrometers, 0.02 micrometers, 0.1 micrometers, 0.2 micrometers, 0.22 micrometers, 0.45 micrometers, 1. micrometer, 1.1 micrometers, 2.5 micrometers, 6. micrometers, 20. micrometers, 30. micrometers, or 100. micrometers or Null.
PrefilterPoreSize
The pore size of the filter; all particles larger than this should be removed during the filtration.
Pattern Description: 0.008 micrometers, 0.02 micrometers, 0.1 micrometers, 0.2 micrometers, 0.22 micrometers, 0.45 micrometers, 1. micrometer, 1.1 micrometers, 2.5 micrometers, 6. micrometers, 20. micrometers, 30. micrometers, or 100. micrometers or Null.
FilterSyringe
Default Calculation: Resolves to an syringe appropriate to the volume of sample being filtered, if Filtration is set to True.
Pattern Description: An object of type or subtype Model[Container, Syringe] or Object[Container, Syringe] or a prepared sample or Null.
Programmatic Pattern: ((ObjectP[{Model[Container, Syringe], Object[Container, Syringe]}] | _String) | Automatic) | Null
FilterHousing
The filter housing that should be used to hold the filter membrane when filtration is performed using a standalone filter membrane.
Default Calculation: Resolve to an housing capable of holding the size of the membrane being used, if filter with Membrane FilterType is being used and Filtration is set to True.
Pattern Description: An object of type or subtype Model[Instrument, FilterHousing], Object[Instrument, FilterHousing], Model[Instrument, FilterBlock], or Object[Instrument, FilterBlock] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, FilterHousing], Object[Instrument, FilterHousing], Model[Instrument, FilterBlock], Object[Instrument, FilterBlock]}] | Automatic) | Null
FilterIntensity
Default Calculation: Will automatically resolve to 2000 GravitationalAcceleration if FiltrationType is Centrifuge and Filtration is True.
Pattern Description: Greater than 0 revolutions per minute or greater than 0 standard accelerations due to gravity on the surface of the earth or Null.
Programmatic Pattern: ((GreaterP[0*RPM] | GreaterP[0*GravitationalAcceleration]) | Automatic) | Null
FilterTime
Default Calculation: Will automatically resolve to 5 Minute if FiltrationType is Centrifuge and Filtration is True.
FilterTemperature
The temperature at which the centrifuge chamber will be held while the samples are being centrifuged during filtration.
Default Calculation: Will automatically resolve to 22 Celsius if FiltrationType is Centrifuge and Filtration is True.
FilterContainerOut
The desired container filtered samples should be produced in or transferred into by the end of filtration, with indices indicating grouping of samples in the same plates, if desired.
Default Calculation: Automatically set as the PreferredContainer for the Volume of the sample. For plates, attempts to fill all wells of a single plate with the same model before using another one.
Pattern Description: An object of type or subtype Model[Container] or Object[Container] or a prepared sample or {Index, Container} or Null.
Programmatic Pattern: (((ObjectP[{Model[Container], Object[Container]}] | _String) | {GreaterEqualP[1, 1] | Automatic, (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
FilterAliquotDestinationWell
The desired position in the corresponding AliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
FilterAliquotContainer
The desired type of container that should be used to prepare and house the filter samples which should be used in lieu of the SamplesIn for the experiment.
Programmatic Pattern: ((ObjectP[Model[Container]] | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
FilterAliquot
The amount of each sample that should be transferred from the SamplesIn into the FilterAliquotContainer when performing an aliquot before filtration.
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container.
Pattern Description: All or greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
FilterSterile
Default Calculation: Resolve to False if Filtration is indicated. If sterile filtration is desired, this option must manually be set to True.
Aliquoting
Aliquot
Indicates if aliquots should be taken from the SamplesIn and transferred into new AliquotSamples used in lieu of the SamplesIn for the experiment. Note that if NumberOfReplicates is specified this indicates that the input samples will also be aliquoted that number of times. Note that Aliquoting (if specified) occurs after any Sample Preparation (if specified).
AliquotAmount
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container if a liquid, or the current Mass or Count if a solid or counted item, respectively.
Programmatic Pattern: ((RangeP[1*Microliter, 20*Liter] | RangeP[1*Milligram, 20*Kilogram] | GreaterP[0*Unit, 1*Unit] | GreaterP[0., 1.] | All) | Automatic) | Null
TargetConcentration
The desired final concentration of analyte in the AliquotSamples after dilution of aliquots of SamplesIn with the ConcentratedBuffer and BufferDiluent which should be used in lieu of the SamplesIn for the experiment.
TargetConcentrationAnalyte
Default Calculation: Automatically set to the first value in the Analytes field of the input sample, or, if not populated, to the first analyte in the Composition field of the input sample, or if none exist, the first identity model of any kind in the Composition field.
Pattern Description: An object of type or subtype Model[Molecule], Model[Molecule, cDNA], Model[Molecule, Oligomer], Model[Molecule, Transcript], Model[Molecule, Protein], Model[Molecule, Protein, Antibody], Model[Molecule, Carbohydrate], Model[Molecule, Polymer], Model[Resin], Model[Resin, SolidPhaseSupport], Model[Lysate], Model[ProprietaryFormulation], Model[Virus], Model[Cell], Model[Cell, Mammalian], Model[Cell, Bacteria], Model[Cell, Yeast], Model[Tissue], Model[Material], or Model[Species] or Null.
AssayVolume
Default Calculation: Automatically determined based on Volume and TargetConcentration option values.
Pattern Description: Greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
ConcentratedBuffer
The concentrated buffer which should be diluted by the BufferDilutionFactor in the final solution (i.e., the combination of the sample, ConcentratedBuffer, and BufferDiluent). The ConcentratedBuffer and BufferDiluent will be combined and then mixed with the sample, where the combined volume of these buffers is the difference between the AliquotAmount and the total AssayVolume.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BufferDilutionFactor
The dilution factor by which the concentrated buffer should be diluted in the final solution (i.e., the combination of the sample, ConcentratedBuffer, and BufferDiluent). The ConcentratedBuffer and BufferDiluent will be combined and then mixed with the sample, where the combined volume of these buffers is the difference between the AliquotAmount and the total AssayVolume.
Default Calculation: If ConcentratedBuffer is specified, automatically set to the ConcentratedBufferDilutionFactor of that sample; otherwise, set to Null.
BufferDiluent
The buffer used to dilute the aliquot sample such that ConcentratedBuffer is diluted by BufferDilutionFactor in the final solution. The ConcentratedBuffer and BufferDiluent will be combined and then mixed with the sample, where the combined volume of these buffers is the difference between the AliquotAmount and the total AssayVolume.
Default Calculation: Automatically resolves to Model[Sample, "Milli-Q water"] if ConcentratedBuffer is specified; otherwise, resolves to Null.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
AssayBuffer
The buffer that should be added to any aliquots requiring dilution, where the volume of this buffer added is the difference between the AliquotAmount and the total AssayVolume.
Default Calculation: Automatically resolves to Model[Sample, "Milli-Q water"] if ConcentratedBuffer is not specified; otherwise, resolves to Null.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
AliquotSampleStorageCondition
The non-default conditions under which any aliquot samples generated by this experiment should be stored after the protocol is completed.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting, Oven} or Disposal or Null.
DestinationWell
The desired position in the corresponding AliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
Pattern Description: Any well from A1 to H12 or list of one or more any well from A1 to H12 or any well from A1 to H12 entries or Null.
Programmatic Pattern: ((WellPositionP | {((Automatic | Null) | WellPositionP)..}) | Automatic) | Null
AliquotContainer
The desired type of container that should be used to prepare and house the aliquot samples, with indices indicating grouping of samples in the same plates, if desired. This option will resolve to be the length of the SamplesIn * NumberOfReplicates.
Default Calculation: Automatically set as the PreferredContainer for the AssayVolume of the sample. For plates, attempts to fill all wells of a single plate with the same model before aliquoting into the next.
Pattern Description: An object of type or subtype Model[Container] or Object[Container] or a prepared sample or Automatic or Null or {Index, Container} or list of one or more an object of type or subtype Model[Container] or Object[Container] or a prepared sample or Automatic or Null entries or list of one or more Automatic or Null or {Index, Container} entries.
Programmatic Pattern: (((ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null) | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null)} | {((ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null))..} | {({GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null)} | (Automatic | Null))..}) | Automatic) | Null
AliquotPreparation
Default Calculation: Automatic resolution will occur based on manipulation volumes and container types.
ConsolidateAliquots
Protocol Options
Organizational Information
Template
A template protocol whose methodology should be reproduced in running this experiment. Option values will be inherited from the template protocol, but can be individually overridden by directly specifying values for those options to this Experiment function.
Pattern Description: An object of type or subtype Object[Protocol] or an object of type or subtype of Object[Protocol] with UnresolvedOptions, ResolvedOptions specified or Null.
Programmatic Pattern: (ObjectP[Object[Protocol]] | FieldReferenceP[Object[Protocol], {UnresolvedOptions, ResolvedOptions}]) | Null
Name
A object name which should be used to refer to the output object in lieu of an automatically generated ID number.
Post Experiment
MeasureWeight
Indicates if any solid samples that are modified in the course of the experiment should have their weights measured and updated after running the experiment. Please note that public samples are weighed regardless of the value of this option.
MeasureVolume
Indicates if any liquid samples that are modified in the course of the experiment should have their volumes measured and updated after running the experiment. Please note that public samples are volume measured regardless of the value of this option.
ImageSample
Example Calls
CartridgeType
Detects the concentrations of one analyte (e.g., Interleukin 6) in the samples using a pre-loaded SinglePlex72X1 capillary ELISA cartridge:
Detects the concentrations of multiple analytes in the samples simultaneously using a multi-analyte capillary ELISA cartridge. For the current list of available capillary ELISA cartridges, please perform Search[Model[Container,Plate,Irregular,CapillaryELISA]] with the desired CartridgeType, AnalyteMolecules or other conditions:
Customized sandwich ELISA assays can be performed on the samples using a Customizable capillary ELISA cartridge:
A new pre-loaded capillary ELISA cartridge can be created and ordered by setting Cartridge to Null and choose the desired values in CartridgeType, Species and Analytes options:
Sample Spiking
The input samples can be mixed with the desired spike samples with known concentration(s) of analyte(s) to increase the analytical response of the unknown samples, for evaluation of sample matrices in a new ELISA assay:
Sample Dilution
A dilution curve can be specified for an input sample by providing the volumes of the diluted samples and the dilution factor at each point of the curve. All the diluted samples are loaded in the capillary ELISA cartridge:
A stepwise serial dilution curve can be specified for an input sample by providing the volume of the samples, a serial dilution factor and the number of times the sample should be diluted. All the diluted samples are loaded in the capillary ELISA cartridge. This should be used if you want the sample to be diluted by the same factor at each step of the serial dilution. The volume of the sample will be the final volume of the sample after the dilution is performed. The example call below will create a series of samples with dilution factors of (0.5, 0.25, 0.13, 0.063, 0.031, 0.016, 0.0078, 0.0039) and a volume of 60 Microliters:
A stepwise serial dilution curve can be specified for an input sample by providing the volume of the samples and a series of dilution factors the sample should be diluted by at each step. All the diluted samples are loaded in the capillary ELISA cartridge. This should be used if you want the sample to be diluted by different factors at each step of the serial dilution. The volume of the sample will be the final volume of the sample after the dilution is performed. The example call below will create a series of samples with dilution factors of (0.5, 0.05, 0.01) and a volume of 60 Microliters:
A stepwise serial dilution curve can be specified for an input sample by providing the volume of the sample that should be transferred, the volume of diluent the sample should be added to and the number of times the sample should be diluted. All the diluted samples are loaded in the capillary ELISA cartridge. This should be used if you want give the volumes the sample will be diluted by with instead of dilution factors. The example call below will create a series of samples with dilution factors of (0.67, 0.44, 0.30, 0.20, 0.13, 0.09) and a volume of 50 Microliters:
The desired diluent can be used to dilute the input samples before they are loaded into the capillary ELISA cartridge:
Standard
Standard samples can be added to the capillary ELISA experiment to provide information about the accuracy of the assay or be used for the generation of the standard curves:
A stepwise serial dilution curve of the standard can be specified for an input sample by providing the volume of the standard, a serial dilution factor and the number of times the sample should be diluted:
The solid-state Standard samples can be resuspended to the desired concentrations in solution using the desired diluents before dilution or cartridge loading and can be stored for future use:
Antibody
Capture antibody and detection antibody samples can be specified for samples in a Customizable capillary ELISA cartridge
Antibody samples can be resuspended if they are in solid state following the instructions provided in the corresponding options:
Warnings and Errors
Messages (175)
CannotCreateNewCustomizableCartridge (1)
CannotSpecifyAntibodyOptions (1)
CannotSpecifyCaptureAntibodyConjugationOptions (1)
CannotSpecifyCaptureAntibodyDilutionOptions (1)
CannotSpecifyCaptureAntibodyResuspensionOptions (1)
CannotSpecifyDetectionAntibodyConjugationOptions (1)
CannotSpecifyDetectionAntibodyDilutionOptions (1)
CannotSpecifyDetectionAntibodyResuspensionOptions (1)
CannotSpecifySpikeOptions (1)
CannotSpecifyStandardAntibodyOptions (1)
CannotSpecifyStandardCaptureAntibodyConjugationOptions (1)
CannotSpecifyStandardCaptureAntibodyDilutionOptions (1)
CannotSpecifyStandardCaptureAntibodyResuspensionOptions (1)
CannotSpecifyStandardDetectionAntibodyConjugationOptions (1)
CannotSpecifyStandardDetectionAntibodyDilutionOptions (1)
CannotSpecifyStandardDetectionAntibodyResuspensionOptions (1)
CannotSpecifyStandardOptions (1)
CannotSpecifyStandardResuspensionOptions (1)
CannotUseMultiPlex32X8CartridgeForNonHuman (1)
CapillaryELISAConflictingStorageConditions (1)
CapillaryELISASampleDilution (1)
CapillaryELISAStandardDilution (1)
CaptureAntibodyDilutionRecommended (1)
ConflictAntibodyEpitopes (1)
ConflictCapillaryELISAAntibodyObjectSample (1)
ConflictCapillaryELISAAntibodyOptions (1)
ConflictCapillaryELISAStandardComposition (1)
ConflictCaptureAntibodyConjugationOptions (1)
ConflictCaptureAntibodyDilutionOptions (1)
ConflictCartridgeAndAnalytes (2)
ConflictCartridgeAndCartridgeType (1)
ConflictDetectionAntibodyConjugationOptions (1)
ConflictDetectionAntibodyDilutionOptions (1)
ConflictPreLoadedAnalytes (1)
ConflictStandardAntibodyEpitopes (1)
ConflictStandardCaptureAntibodyConjugationOptions (1)
ConflictStandardCaptureAntibodyDilutionOptions (1)
ConflictStandardDetectionAntibodyConjugationOptions (1)
ConflictStandardDetectionAntibodyDilutionOptions (1)
ConflictStandardDilutionCurve (1)
DeprecatedCapillaryELISAInstrumentModel (1)
DetectionAntibodyDilutionRecommended (1)
DiscardedCapillaryELISACartridge (1)
DuplicatedAnalytes (2)
DuplicateName (1)
EmptyCartridgeChannel (1)
ExceedCapillaryELISACartridgeCapacity (2)
The total number of loading samples including all diluted samples must be smaller than or equal to the capacity of the capillary ELISA cartridge (48 for a customizable cartridge):
The total number of loading samples including all diluted samples must be smaller than or equal to the capacity of the capillary ELISA cartridge:
ExceedCaptureAntibodyConjugationContainerCapacity (1)
ExceedCaptureAntibodyPurificationColumnCapacity (1)
ExceedDetectionAntibodyConjugationContainerCapacity (1)
ExceedDetectionAntibodyPurificationColumnCapacity (1)
ExceedLengthAnalytesForCustomizableCartridge (1)
ExceedStandardCaptureAntibodyConjugationContainerCapacity (1)
ExceedStandardCaptureAntibodyPurificationColumnCapacity (1)
ExceedStandardComposition (1)
ExceedStandardDetectionAntibodyConjugationContainerCapacity (1)
ExceedStandardDetectionAntibodyPurificationColumnCapacity (1)
IncompleteResolvedSpikeConcentration (1)
IncompleteResolvedStandardComposition (1)
InvalidCaptureAntibodyResuspensionForLiquid (1)
InvalidCaptureAntibodyResuspensionForSolid (1)
InvalidDetectionAntibodyResuspensionForLiquid (1)
InvalidDetectionAntibodyResuspensionForSolid (1)
InvalidStandardCaptureAntibodyResuspensionForLiquid (1)
InvalidStandardCaptureAntibodyResuspensionForSolid (1)
InvalidStandardDetectionAntibodyResuspensionForLiquid (1)
InvalidStandardDetectionAntibodyResuspensionForSolid (1)
InvalidStandardOptionLength (1)
InvalidStandardResuspensionForLiquid (1)
InvalidStandardResuspensionForSolid (1)
LongLeadTimeCartridge (1)
MustSpecifyBiotinReagentVolume (1)
MustSpecifyCaptureAntibody (1)
MustSpecifyCaptureAntibodyConjugationOptions (1)
MustSpecifyCaptureAntibodyDilutionOptions (1)
MustSpecifyCaptureAntibodyLoadingVolume (1)
MustSpecifyCaptureAntibodyResuspensionOptions (1)
MustSpecifyDetectionAntibody (1)
MustSpecifyDetectionAntibodyConjugationOptions (1)
MustSpecifyDetectionAntibodyDilutionOptions (1)
MustSpecifyDetectionAntibodyLoadingVolume (2)
MustSpecifyDetectionAntibodyResuspensionOptions (1)
MustSpecifyDigoxigeninReagentVolume (1)
MustSpecifyPreLoadedCartridgeAnalytes (1)
MustSpecifySpikeOptions (1)
MustSpecifyStandardBiotinReagentVolume (1)
MustSpecifyStandardCaptureAntibody (1)
MustSpecifyStandardCaptureAntibodyConjugationOptions (1)
MustSpecifyStandardCaptureAntibodyDilutionOptions (1)
MustSpecifyStandardCaptureAntibodyLoadingVolume (1)
MustSpecifyStandardCaptureAntibodyResuspensionOptions (1)
MustSpecifyStandardDetectionAntibody (1)
MustSpecifyStandardDetectionAntibodyConjugationOptions (1)
MustSpecifyStandardDetectionAntibodyDilutionOptions (1)
MustSpecifyStandardDetectionAntibodyLoadingVolume (2)
MustSpecifyStandardDetectionAntibodyResuspensionOptions (1)
MustSpecifyStandardDigoxigeninReagentVolume (1)
MustSpecifyStandardOptions (1)
MustSpecifyStandardResuspensionOptions (1)
NoCommonDiluentForAnalytes (1)
NoCommonMinDilutionFactorForAnalytes (1)
NoELISAAssayTypeForAntibodySamples (2)
NonOptimalCapillaryELISASampleDilution (1)
NonOptimalCaptureAntibodyDiluent (1)
NonOptimalCaptureAntibodyDilution (1)
NonOptimalCaptureAntibodyPurificationColumn (1)
NonOptimalDetectionAntibodyDiluent (1)
NonOptimalDetectionAntibodyDilution (1)
NonOptimalDetectionAntibodyPurificationColumn (1)
NonOptimalLoadingVolume (1)
NonOptimalStandardCaptureAntibodyDiluent (1)
NonOptimalStandardCaptureAntibodyDilution (1)
NonOptimalStandardCaptureAntibodyPurificationColumn (1)
NonOptimalStandardDetectionAntibodyDiluent (1)
NonOptimalStandardDetectionAntibodyDilution (1)
NonOptimalStandardDetectionAntibodyPurificationColumn (1)
NonOptimalStandardDiluent (1)
NonOptimalStandardDilutionCurve (1)
NonOptimalStandardLoadingVolume (1)
NonOptimalStandardResuspensionConcentration (1)
NonOptimalWashBuffer (1)
NotEnoughBiotinReagentVolume (1)
NotEnoughDigoxigeninReagentVolume (1)
NotEnoughDilutionPreparationVolume (1)
NotEnoughSampleVolumeAndSpikeVolume (4)
Enough volume of input samples must be provided to make the desired dilutions (defaulted to 30 Microliter required volume for DilutionCurve):
Enough volume of input samples must be provided to make the desired dilutions:
Enough volume of input samples plus spike samples must be provided to make the desired dilutions:
Enough volume of input samples plus spike samples (if applicable) must be provided to make the desired dilutions (defaulted to 30 Microliter required volume for DilutionCurve):
NotEnoughStandardBiotinReagentVolume (1)
NotEnoughStandardDigoxigeninReagentVolume (1)
NotEnoughStandardDilutionPreparationVolume (1)
ObjectDoesNotExist (6)
Throw a message if we have a sample that does not exist (name form):
Throw a message if we have a container that does not exist (name form):
Throw a message if we have a sample that does not exist (ID form):
Throw a message if we have a container that does not exist (ID form):
Do NOT throw a message if we have a simulated sample but a simulation is specified that indicates that it is simulated:
Do NOT throw a message if we have a simulated container but a simulation is specified that indicates that it is simulated:
RetiredCapillaryELISAInstrument (1)
StandardCaptureAntibodyDilutionRecommended (1)
StandardDetectionAntibodyDilutionRecommended (1)
TooHighCaptureAntibodyDilution (1)
TooHighDetectionAntibodyDilution (1)
TooHighStandardCaptureAntibodyDilution (1)
TooHighStandardDetectionAntibodyDilution (1)
TooLargeDilutionMixVolumeForCapillaryELISASample (1)
TooLargeDilutionMixVolumeForCapillaryELISAStandard (1)
TooManyAnalytes (2)
TooManyAnalytesForSampleNumber (1)
TooManyLoadingSamplesForCapillaryELISA (1)
UnsupportedAnalytes (1)
UnsupportedCaptureAntibodyPurificationColumn (1)
UnsupportedDetectionAntibodyPurificationColumn (1)
UnsupportedStandardCaptureAntibodyPurificationColumn (1)
UnsupportedStandardDetectionAntibodyPurificationColumn (1)
WrongAnalyteSpikeConcentration (1)
WrongAnalyteStandardComposition (1)
WrongCartridgeTypeAnalytes (3)
Possible Issues
Insufficient Loading Volumes
For the best ELISA performance, it is recommended to keep the sample loading volumes as 50 Microliter or more. Lower cartridge loading volumes may lead to inaccurate measurements of fluorescence intensities and analyte concentrations.
Insufficient Dilution
The samples should be diluted more than the recommended minimum dilution factor determined by the assay developer for the best performance of the ELISA assays. Insufficient dilution may lead to higher background intensities and fluorescence intensities that are out of the detector's detection range or the linear quantitation range of the assay. For the minimum dilution factors of different ELISA assays, please inspect the Object[ManufacturingSpecification,CapillaryELISACartridge]s of different analytes.
Non-optimal Diluent
The samples should be diluted using the recommended sample diluent provided by the assay developer for the best performance of the ELISA assays. Non-optimal sample diluents may lead to higher background intensities and unexpected microfluidic behaviors. For the recommended diluents of different ELISA assays, please inspect the Object[ManufacturingSpecification,CapillaryELISACartridge]s of different analytes.
Non-optimal Wash Buffer
The wash buffer of the capillary ELISA cartridge should be kept as Model[Sample,"Simple Plex Wash Buffer"] provided by the assay developer for the best performance of the ELISA assays. Non-optimal sample diluent may lead to higher background intensities and unexpected microfluidic behaviors.
Non-optimal Customizable Assays
It is recommended to perform a series of ELISA assays on customizable capillary ELISA cartridges using the desired custom antibody samples in order to select the best assay conditions and parameters for resuspension, conjugation and dilution processes. For the analytes with validated pre-loaded assays available, it is highly recommended to use pre-loaded capillary ELISA cartridges for the best results.
Last modified on Fri 15 Aug 2025 17:21:07