Av. Zherdev et al., INTERACTION BETWEEN ANTIBODIES AND HAPTEN-PROTEIN CONJUGATES OF DIFFERENT COMPOSITION - THEORETICAL PREDICTIONS AND EXPERIMENTAL-DATA, Journal of immunoassay, 18(1), 1997, pp. 67-95
Mathematical models of competitive ELISAs with labelled antibody and w
ith labelled antigen taking into account bivalent interactions between
antibodies and hapten-protein conjugates were developed and analysed.
It was shown that in the kinetic model of the immunochemical reaction
the conjugate composition influenced the amplitude of detected signal
but not ELISA sensitivity. In the equilibrium model decreased sensiti
vity correlated with bivalent complexes formation. The predictions wer
e tested experimentally using 2,4-dichlorophenoxyacetic acid (2,4-D) a
nd testosterone as haptens, It was confirmed that increasing of the ha
pten : protein ratio resulted in formation of bivalent complexes with
antibodies. The equilibrium binding constants for these complexes were
two orders of magnitude higher than for monovalent ones. Optimal conj
ugate compositions have been chosen for ELISA of these haptens.