ENZYME-LINKED IMMUNOSORBENT ASSAYS IN A CHROMATOGRAPHIC FORMAT

This paper describes the execution of enzyme-linked immunosorbent assa ys (ELISA), (i) in immunosorbent columns with antibodies immobilized o n porous particles, (ii) where samples and reagents are metered by val ves and syringe pumps, (iii) samples, reagents, substrate, and wash bu ffers are transported into or through the system by high pressure liqu id chromatography pumps, and (iv) enzyme reaction product is detected by absorbance. The normal protocol used for ELISA was modified in that antigen was complexed with enzyme conjugated antibody in the autosamp ler and an aliquot introduced into the system where it was transported to the immunosorbent and captured. Substrate was subsequently pumped into the immunosorbent bed and the product swept to an absorbance dete ctor for quantitation.