Factors influencing DTPA conjugation with antibodies by cyclic DTPA anhydride.
Diethylenetriaminepentaacetic acid (DTPA) was conjugated with a practical concentration (300 micrograms/ml) of antibody to human albumin (Ab) and 1083 17-1A monoclonal colorectal antibody (MAb-17-1A) via an acylation reaction using cyclic DTPA anhydride (cDTPAA). The conjugation reaction was favored as pH increased. Bicarbonate buffer at pH 8.2 was chosen for studies of the effect of the cDTPAA-to-antibody ratio on DTPA conjugation with antibody because of its good buffer capacity at that pH. The reaction of cDTPAA with Ab at molar ratios of 2000, 1000, 500, and 100 in the bicarbonate buffer gave rise to 11, 9, 8, and 2 indium atoms incorporated per Ab with 47%, 55%, 59%, and 77% retention of the binding activity. For the conjugation reaction of MAb-17-1A, 29, 28, 31, 11, 4, and 1 indium atoms were incorporated, with the retention of less than 5%, less than 5%, less than 5%, 12%, 60%, and 93% of binding activity when the molar ratio was 5000, 2000, 1000, 500, 100, and 50.
D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D055598:Chemical Phenomena; D002621:Chemistry; D003110:Colonic Neoplasms / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D004369:Pentetic Acid / Q000031:analogs & derivatives*; D011817:Rabbits; D012004:Rectal Neoplasms / Q000276:immunology; D012709:Serum Albumin / Q000276:immunology
D000906:Antibodies; D000911:Antibodies, Monoclonal; D012709:Serum Albumin; C039777:cyclic DTPA anhydride; D004369:Pentetic Acid