Hu R, Xie G Y, Zhang X, Guo Z Q, Jin S
College of Chemistry and Molecular Engineering, Peking University, Beijing, China.
J Biotechnol. 1998 Apr 15;61(2):109-15. doi: 10.1016/s0168-1656(98)00009-1.
A new approach for producing catalytic anti-idiotypic antibody was developed. A monoclonal anti-idiotypic antibody which was induced against carboxypeptidase A (CPA) showed the catalytic activity similar to the original antigen. The activity of the catalytic antibody was investigated. Rabbits were immunized by bovine pancreas carboxypeptidase A. The antiserum was purified and used as antigen to immunize BALB/c mice to induce monoclonal anti-idiotypic antibodies. Screened for enzymatic activities, the monoclonal antibody 32C3 showed esterase activity. The hydrolysis of hippuryl-DL-phenyllactic acid by McAb 32C3 followed the enzymatic kinetics. In our experimental system, Kcat value was 0.0123 min-1 and Km was 0.04M. The acceleration rate was 1750 times compared to the rate of self-hydrolysis of the substrate. This hydrolysis reaction can be competitively inhibited by hydrocinnamic acid. This method could be effective to obtain catalytic antibodies with the characters close to natural enzymes.
开发了一种生产催化抗独特型抗体的新方法。一种针对羧肽酶A（CPA）诱导产生的单克隆抗独特型抗体表现出与原始抗原相似的催化活性。对催化抗体的活性进行了研究。用牛胰腺羧肽酶A免疫兔子。纯化抗血清并用作抗原免疫BALB/c小鼠以诱导单克隆抗独特型抗体。经酶活性筛选，单克隆抗体32C3表现出酯酶活性。McAb 32C3对马尿酸-DL-苯乳酸的水解遵循酶促动力学。在我们的实验系统中，Kcat值为0.0123 min-1，Km为0.04M。与底物的自水解速率相比，加速率为1750倍。该水解反应可被氢化肉桂酸竞争性抑制。该方法可能有效地获得具有接近天然酶特性的催化抗体。
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