Monoclonal Antibody Testing and Screening
In addition to standard ELISA testing of custom monoclonal antibody cell lines, we offer a variety of additional screening and assay testing service options to validate monoclonal antibodies for use in specific applications.
Screening options for monoclonal antibody development
Our standard monoclonal antibody development service includes ELISA testing to determine which mice (or rats) to use for fusion and ELISA screening of the resulting hybridomas to determine the best cell lines to deliver. Options for more specialized screening and testing are described here. The listed assay strategies demonstrate our broad capabilities to provide nearly any sort of screening and assay validation that customers require for antibody production and purification.
If a quantitative immunoassay is needed, we can develop suitable matched pairs for your ELISA assay. We utilize the expertise of our own internal ELISA development team which supports more than 50 standard cytokine and chemokine ELISA products. Projects can be initiated in conjunction with our custom hybridoma development services or antibodies can be supplied by customers for evaluation.
Supernatants from positive fusions are screened with the Biacore by injecting supernatant at a concentration of approximately 0.1 -0.25 mg/mL over an anti-mouse IgG Fc surface, followed by injection of the recombinant antigen.
Strong binding candidates are purified via Protein G or via 96-well melon gel depending on number of positive clones (see antibody purification kits and products).
Antibody testing | |
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Plates are coated with purified antibody/s. |
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ELISA assay/s are run to determine best antibody pair. |
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ELISA assay/s is/are run to confirm activity with natural samples. Obtain natural samples from client. |
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The following antibody testing options are available for IHC/IF through our comprehensive antibody development platform. A final report that includes pathological scoring notes for specificity and intensity for tissues or fluorescent staining images and report for IF will be provided.
Qualification testing
Antibodies will be tested against researcher-supplied tissues or cell lines under standard tissue preparatory conditions for formulin-fixed, paraffin-embedded samples for tissues or with standard Thermo Scientific Cellomics Testing for IF.
Antibody testing | |
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Confirm antibody reactivity from customer and expected expression patterns |
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Obtain a minimum of two positive control and one negative control tissues / cell lines from customer |
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Testing antibodies |
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Data |
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The following western blotting testing options is available for any antibody developed. Standard conditions for gel gradient and testing samples will apply. Detection will be monitored by Pierce Supersignal chemiluminescence.
Qualification testing
Antibodies will be tested against researcher-supplied tissues or cell lines under standard preparatory conditions.
Simple western blot testing
Confirm antibody reactivity from customer and expected expression patterns by source and molecular weight.
- Expected tissue or cellular distribution, including overexpression construct and negative control if available (3 samples maximum)
- Electrophoresis by appropriate tris-glycine gradient gel to optimize for predicted or confirmed molecular weight.
- Antibodies will be tested at 3 working dilutions (1:100, 1:200, 1:500) unless otherwise directed.
Advanced western testing
Antibodies are tested in the same manner as described for "Simple Western blot testing" except samples are enriched for specific cellular locations prior to phoresis and up to 5 samples may be tested for each antibody. This is an important offering for customers who know their protein(s) of interest may be abundant at lower concentrations.
Confirm antibody reactivity from customer and expected expression patterns by source and molecular weight.
- Expected tissue or cellular distribution, including overexpression construct and negative control if available (5 samples maximum).
- Prepared subcellular fractions using appropriate subcellular fractionation kit.
- Electrophoresis by appropriate tris-glycine gradient gel to optimize for predicted or confirmed molecular weight.
- Antibodies will be tested at 3 working dilutions (1:100, 1:200, 1:500) unless otherwise directed.
Tools and information
Support
For Research Use Only. Not for use in diagnostic procedures.