Determination of a monoclonal antibody binding activity using immunodetection.
A method for measuring the binding of chimeric IgG BR-96 to the Lewis Y antigen using the ImmunoDetection technology has been developed. The procedure is rapid (2 min), highly reproducible (< 5% CV), and has excellent correlation with the Lewis Y and anti-idiotypic enzyme immunoassays. Samples that were degraded by heat or by repeated freeze-thaw cycles showed reduced binding activity. There was minimal cross-reactivity with other proteins typically found in hybridoma media or with another chimeric IgG directed against a different antigen (L6). The fact that this assay can be performed using conventional HPLC equipment makes it especially attractive because it can be completely automated using equipment readily available.
Schenerman
M A
MA
Biotechnology Development Laboratories, Bristol-Myers Squibb Company, Syracuse, New York 13221.
Collins
T J
TJ
D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D002248:Carbon Monoxide / Q000378:metabolism; D005240:Feasibility Studies; D007074:Immunoglobulin G / Q000378:metabolism; D007158:Immunologic Techniques; D007700:Kinetics; D007983:Lewis Blood Group Antigens; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007983:Lewis Blood Group Antigens; D011993:Recombinant Fusion Proteins; D002248:Carbon Monoxide