The majority of hybridoma projects use indirect ELISA to screen hybridomas after HAT selection. However, ELISA-guaranteed antibody generally refers to two antibodies that can be used in a sandwich ELISA or two antibodies that bind different epitopes. Selecting a pair of capture antibodies and detection antibodies is very technical. Only pairs with high specificity, selectivity, and consistent linearity of dilution for the target will be used for further development. Plasma, serum, or tissue lysate will be used to validate antibody selectivity. In many cases, purified recombinant target proteins will be added into the biological matrix, which should be recovered and detected. The recovery observed for the spike should be almost identical in both the biological matrix and the standard diluent for a sample matrix to be considered valid for our ELISA assay. The development of LFIA functional antibodies generally involves the use of a chromatography platform to verify the function of the antibody after hybridoma is subcloned and cryopreserved and antibody is purified.

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