Study on site-directed coupling method of immunoaffinity Chromatography

Huang Xin; et al

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Study on site-directed coupling method of immunoaffinity Chromatography

更新时间：2024-01-17

- Study on site-directed coupling method of immunoaffinity Chromatography
- Journal of Chongqing Medical University Issue 2, (1999)
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  CLC： R392-33
- Published：1999，
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[1]黄昕,邱宗荫.免疫亲和色谱定向偶联方法的研究[J].重庆医科大学学报,1999(02):132-134.

Huang Xin, et al. Study on site-directed coupling method of immunoaffinity Chromatography. [J]. Journal of Chongqing Medical University (2).(1999)
[1]黄昕,邱宗荫.免疫亲和色谱定向偶联方法的研究[J].重庆医科大学学报,1999(02):132-134. DOI：

Huang Xin, et al. Study on site-directed coupling method of immunoaffinity Chromatography. [J]. Journal of Chongqing Medical University (2).(1999) DOI：

摘要

将初步纯化属小鼠ＩｇＧ１亚类的抗Ｇ３单克隆抗体（ＭＧ３１－随机偶联于ＣＮＢｒ－Ｓｅｐｈａｒｏｓｅ４日凝胶，并成功地定向偶联于ｒＰｒｏｔｅｉｎＡ－ＳｅｐｈａｒｏｓｅＦＦ凝胶，两种偶联反应的结合率分别达到９１．７％和８６．６％。ＭＧ３１－ｒＰｒｏｔｅｉｎＡＳｅｐｈａｒｏｓｅＦＦ的抗原结合容量是ＭＧ３１－ＣＮＢｒ－Ｓｅｐｂａｒｏｓｅ４Ｂ的１．８倍，因此，单克隆抗体通过其Ｆｃ区与ｒＰｒｏｔｅｉｎＡ结合，留出正确定向的抗原结合位点，能够提高免疫亲和吸附剂的结合容量。

Abstract

e coupled anti - G3 McAb (MG31

IgG1) purified by ammonium sulphate precipitation method to CNBr - Sepharose 4B gel at random

and to rProtein A - Sepharose FF gel by site - directed immobilization technique successfully. The binding rate of two coupling reaction were 91.7% and 86.6% respectively. The antigen binding capacity of MGd31 - rProtein A - Sepharose FF was 1.8 fold that of MG2-CNBr - 8epharose 4B

thus the binding of McAbs via their Fe portions to rProtein A

leaving the antigen - combining site in thecorrect orientation for the binding of antigen

could increase the antigen binding capacity of the immunoaffinity chromatography column

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