Abstract
<p>A complete protocol for performing sandwich ELISA using pre-coated microplate kits for quantitative detection of target analytes in serum, plasma, cell culture supernatant, and other biological samples. Protocol developed by abinScience / AtaGenix Technical Team. Sandwich ELISA (enzyme-linked immunosorbent assay) uses a matched antibody pair — a capture antibody pre-coated on the microplate and a detection antibody conjugated to biotin or HRP — to quantify a specific antigen. This protocol covers reagent preparation, sample dilution, standard curve generation, plate washing, substrate development, and data analysis. It is optimized for pre-coated 96-well ELISA kits but can be adapted to custom-coated assays. Typical applications include pharmacokinetic (PK) studies, bioprocess titer measurement, and immunogenicity (anti-drug antibody) screening.</p>