Purification of monoclonal antibodies to human alkaline phosphatases by antigen-immunoaffinity chromatography: comparisons of their molar binding values.

ABSTRACT

Antigen-immunoaffinity chromatography has been used to purify monoclonal antibodies to human alkaline phosphatases. The binding of the purified antibodies to various alkaline phosphatases was tested utilizing an electrophoretic titration assay. The results indicate that binding differences among the various ALPs seen with several of the antibodies are due to antigenic differences in the molecular structure of the enzymes. We define a molar binding value as the minimum molar concentration of antibody necessary to complex all of the ALP in a standard reaction mixture. These values varied widely, indicating marked differences in the avidities of the different antibodies.