Dissociation of monoclonal antibody-antigen complexes: implications for ELISA procedures.
A single step solid phase radioimmunoassay (SS-SPRIA) has been developed for human chorionic gonadotropin (hCG) using monoclonal antibodies (MAb) from culture media adsorbed immunochemically on plastic tubes. The assays have been found to be very simple in terms of operation and do not demand purification of MAbs. Several MAbs which do not show any displacement in liquid phase RIA and ELISA provide a satisfactory SS-SPRIA. Our investigations revealed that the assumption regarding the stability of the primary Mab-Ag complex during incubation and washing steps in ELISAs is not strictly valid for dissociable MAbs. A comparison of different assay systems suggests that the single step SPRIA offers additional advantages over conventionally used multistep ELISA procedures and provides a quantitative probe for the analysis of epitope-paratope interactions.
Venkatesh
N
N
Primate Research Laboratory, Indian Institute of Science, Bangalore, India. nvenkat@hamsadvani.serc.iisc.ernet.in
Murthy
G S
GS
D000911:Antibodies, Monoclonal / Q000276:immunology; D000936:Antigen-Antibody Complex / Q000737:chemistry*; D000937:Antigen-Antibody Reactions; D001667:Binding, Competitive; D006063:Chorionic Gonadotropin / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000737:chemistry; D006801:Humans; D011863:Radioimmunoassay / Q000379:methods*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D006063:Chorionic Gonadotropin; D000939:Epitopes