Alterations of specific and nonspecific binding of monoclonal antibody by introduction of acidic and hydrophobic groups.
AB-6 monoclonal antibody (mAb) against bovine serum albumin (BSA) was coupled with beta-alanine or n-propylamine using a heterobifunctional cross-linking reagent, N-(epsilon-maleimidocaproyloxy)succinimide (EMCS) and a linker, poly-L-cysteine to introduce acidic or hydrophobic groups into the mAb. The mAb molecules coupled with beta-alanine at lower than 1:16 molar ratios and those coupled with n-propylamine at 1:14 retained the original reactivity to BSA but modification with higher amounts of beta-alanine or n-propylamine decreased the reactivity. In contrast, nonspecific bindings of AB-6 mAb to a plastic culture plate and tumor (MDA-MB-453) cells were decreased by modification with beta-alanine and increased by n-propylamine. These results suggested that introduction of acidic groups, and hydrophobic groups to antibody leads to fluctuations in the non-specific binding of antibody.
Department of Hygienic Chemistry, Pharmaceutical Institute, Tohoku University, Sendai, Japan.
J
J
Wang
T
T
Masuko
H
H
Ueno
Y
Y
Hashimoto
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology*; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D001943:Breast Neoplasms / Q000276:immunology; D002417:Cattle; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D007525:Isoelectric Focusing; D011437:Propylamines / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000276:immunology*; D014407:Tumor Cells, Cultured; D015091:beta-Alanine / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D011437:Propylamines; D015091:beta-Alanine; D012710:Serum Albumin, Bovine; C035263:2-propylamine