Mechanistic and Experimental Approaches to Assay Design and Associated Issues with Performance Verification
*Beimar N Iriarte, Abbott Laboratories 

Keywords: assay design mechanistic experimental

The classical approach to an immunoassay design is that of reagent optimization. The process starts with the identification of the marker or analyte, followed by the selection of the assay biological principle, then optimization of the biochemical composition of the buffers for the capture antibody, the conjugate / enzyme / tracer, and the substrate. Concurrently, assay standardization and calibrant matrix composition and concentration levels are established, followed by selection of QC Controls matrix composition and concentration levels. Feasibility evaluation is performed at various stages in this process, thus estimates of precision and accuracy are obtained. At the next stage of development, and in most cases, these data are used for the establishment of pass / fail limits in verification tests, while in others these limits come directly from product requirements. In both cases ris