Measurement of affinity constant of anti-human IgG monoclonal antibodies by an elisa-based method

ABSTRACT

Bakground The affinity of an antibody to its antigen is a crucial parameter in its biological activity and performance of an immunoassay such as ELISA. Affinity of most IgG specific MAbs are often determined by methods which require labeling of either antigen or antibody, and are sometimes difficult to control, do not always lead to the expected signal and often result in immunological modification of the molecules. Moreover, direct solid phase binding assays pose some problems such as diffusion effects and difficulties in reaching equilibrium due to heterogeneous binding and co-operativity