Wang J, Masuko T, Ueno H, Hashimoto Y
Department of Hygienic Chemistry, Pharmaceutical Institute, Tohoku University, Sendai, Japan.
Biol Pharm Bull. 1994 Jun;17(6):831-5. doi: 10.1248/bpb.17.831.
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.
使用异双功能交联剂N-(ε-马来酰亚胺基己酰氧基)琥珀酰亚胺（EMCS）和连接子聚-L-半胱氨酸，将抗牛血清白蛋白（BSA）的AB-6单克隆抗体（mAb）与β-丙氨酸或正丙胺偶联，以将酸性或疏水基团引入该单克隆抗体。以低于1:16的摩尔比与β-丙氨酸偶联的单克隆抗体分子以及以1:14与正丙胺偶联的那些分子保留了对BSA的原始反应性，但用更高量的β-丙氨酸或正丙胺进行修饰会降低反应性。相反，通过用β-丙氨酸修饰可降低AB-6单克隆抗体与塑料培养板和肿瘤（MDA-MB-453）细胞的非特异性结合，而用正丙胺修饰则会增加这种结合。这些结果表明，向抗体中引入酸性基团和疏水基团会导致抗体非特异性结合的波动。
