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Section11:Initial Concept and Method Development of a Competitive Assay
From Assay Guidance Wiki
Competitive Binding Immunoassay
Development and validation of a competition immunoassay requires considerable expertise in reagent characterization and method development. Sandwich and antigen-down immunoassays formats should be explored before attempting the competitive immunoassay format.
Drawbacks Using a Competitive Immunoassay
- A competitive immunoassay is not as sensitive as a sandwich ELISA.
- A competitive immunoassay is more sensitive to matrix issues, especially serum matrix, which can affect assay performance.
- Timing of the various incubation steps is less robust in a competitive assay. That is the IC50 of the standard curve will shift with minor changes in incubation of the various steps of the immunoassay.
- The labeling of the hapten or analyte can change the analyte binding affinity for the antibody. Experiments need to determine the effect of the label on the binding affinity of the antibody to the analyte.
