Ks. Chu et al., STUDIES ON AFFINITY CONSTANTS OF HAPTEN-SPECIFIC MONOCLONAL-ANTIBODIES USING ANTIBODY-IMMOBILIZED ELISA, Biotechnology progress, 11(3), 1995, pp. 352-356
An enzyme-linked immunosorbent assay to measure affinity constants of
hapten-specific monoclonal antibodies has been developed. In this meth
od an antibody-immobilizing format is used. Antibody affinity toward h
apten-enzyme tracer is first calculated by using Scatchard's method. U
sing this affinity, antibody-hapten affinity is then calculated by fol
lowing the principles of chemical thermodynamics and mass conservation
. The affinity from this method agrees with the IC50 values in ELISA.
This method is useful in early determination of affinity during monocl
onal antibody development. Compared with the antigen-coated format, th
is method avoids the problem of distinguishing binding of unliganded a
ntibody and singly liganded antibody to antigens on solid phase.