抑制羧肽酶A催化活性的单克隆抗体的制备及性质测定

北京大学学报（自然科学版）

抑制羧肽酶A催化活性的单克隆抗体的制备及性质测定

胡然¹,余文锋¹,张秀¹,金声¹,郭振泉²

¹北京大学化学与分子工程学院，北京，100871;²北京大学生命科学学院，北京，100871

收稿日期:1995-11-23 出版日期:1996-07-20 发布日期:1996-07-20

The Preparation and Charicterization of the Monoclonal Antibody Inhibiting the Enzymatic Activity of CPA

HU Ran¹, YU Wenfeng¹, ZNANG Xiu¹, JIN Sheng¹, GUO Zhenquan²

¹College of Chemistry and Molecular Engineering, Peking University, 100871; ²College of Life Science, Peking University, 100871

Received:1995-11-23 Online:1996-07-20 Published:1996-07-20

摘要/Abstract

摘要： 以羧肽酶A为抗原，经动物免疫、细胞融合、筛选及3次克隆化培养，获得了一株稳定分泌对羧肽酶A肽酶活性有抑制作用的单克隆抗体的杂交瘤细胞株1G₁₀。染色体数目为80-90；ELISA测定腹水效价为10⁶；亲和常数为1.59×10⁹(mol/L)^-1；免疫球蛋白属IgG₁型；分子量为150 Kd。羧肽酶A与该抗体反应后测定肽酶活性，显示出该抗体能抑制羧肽酶A的肽酶活性，Dixon法作图表明类似竞争性抑制反应，测得其表观抑制常数为1.17×10^-7mol/L。该抗体可能针对羧肽酶A的活性中心部位，为制备具有抗原内影像作用的抗独特型抗体酶打下了基础。

关键词: 单克隆抗体, 羧肽酶A, 抑制酶活性

Abstract: A hybridomas secreting McAb 1G₁₀ which can inhibit the catalytic activity of carboxypeptidase A (CPA) was prepared and characterized. Analyzing the model of cell nucleus, main chromosome number was between 80-90. The titer of the ascites fluid was 10⁶. Binding constant for McAb 1G₁₀ to CPA was 1.59×10⁹(mol/L)^-1. McAb 1G₁₀ was identified as belonging to the IgG₁ subclass.Its molecular weight was about 150 Kd. The interaction of McAb 1G₁₀ with CPA was investigated. A competitive inhibition of peptidase activity of CPA by McAb 1G₁₀ was shown by Dixon Plot. The apparent inhibitory constant obtained by this procedure was 1.17×10^-7 mol/L. These results suggested that the binding site of McAb 1G₁₀ with CPA may be the active site of CPA for its peptidase activity. Based on this work, the idiotypic antibody which is the “Internal-Image” of CPA can be prepared and may have the similar enzymatic activity with CPA.

Key words: monoclonal antibody, CPA, inhibition of enzymatic activity

中图分类号:

胡然,余文锋,张秀,金声,郭振泉 . 抑制羧肽酶A催化活性的单克隆抗体的制备及性质测定[J]. 北京大学学报（自然科学版）.

HU Ran,YU Wenfeng,ZNANG Xiu,JIN Sheng,GUO Zhenquan. The Preparation and Charicterization of the Monoclonal Antibody Inhibiting the Enzymatic Activity of CPA[J]. Acta Scientiarum Naturalium Universitatis Pekinensis.

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https://xbna.pku.edu.cn/CN/Y1996/V32/I4/488

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