Purification of monoclonal antibody against PGRP and its activity for small cell lung cancer in vitro
10.3760/cma.j.issn.1006-9801.2011.07.010
- VernacularTitle:抗前胃泌素释放肽单克隆抗体的纯化及体外对小细胞肺癌的活性
- Author:
Xiaolin ZHOU
;
Zhenwei XUE
;
Zengli LIU
;
Qiaoling XU
;
Meiping CUI
- Publication Type:Journal Article
- Keywords:
Chromatography;
affinity;
Antibodies;
monoclonal;
Purify
- From:
Cancer Research and Clinic
2011;23(7):467-470
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of purification on monoclonal antibody (MAb) against PGRP by Protein A-Sepharose affinity chromatography, and to provide some based data for the purification of other antibody using the same method. Methods The ascites which include MAb was purified by Protein A-Sepharose affinity chromatography. The purity and activity of MAb was tested by SDS-PAGE and ELISA. The biological function was identified by flow cytometer and immunohistochemistry. Results The average concentration of protein in ascites before purification is 23.62 mg/ml. Before and after purification, the total protein is 148.79 mg and 146.67 mg, respectively. The recovery coefficient of protein is 98.58%. The concentration of MAb in ascites is 5.21 mg/ml averagely. The MAb purity is more than 95 %. The immunoactivity of purified antibody is higher than that of unpurified antibody. Conclusion The purity of MAb against PGRP purified by Protein A-Sepharose affinity chromatography is very high. The immunoactivity of purified antibody is higher than that of unpurified antibody. So the ProteinA-Sepharose affinity chromatography is a rapid, convenient and reliable method for the purification of MAb Against PGRP.
