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  

Purification of monoclonal antibody against PGRP and its activity for small cell lung cancer in vitro

作　　者：周小林[1] 薛振伟[1] 刘增礼[2] 徐巧玲[2] 崔梅萍[1] 

机构地区：[1]中国辐射防护研究院放射与环境医学研究所放射医学研究室,太原030006 [2]苏州大学附属第二医院核医学科

出　　处：《肿瘤研究与临床》2011年第7期467-470,共4页Cancer Research and Clinic

基　　金：山西省自然科学基金（2007011128）;中国辐射防护研究院基础研究项目（6160130502）

摘　　要：目的探讨亲和层析法纯化抗前胃泌素释放肽（PGRP）单克隆抗体（MAb）的效果，为该法纯化其他抗体提供数据依据。方法采用蛋白A-琼脂糖亲和层析法纯化含有MAb的腹腔积液，并用SDS—PAGE和酶联免疫吸附（ELISA）法分别检测其纯度和效价，用流式细胞术和免疫组织化学法检测其对小细胞肺癌细胞株NCI—H446的组织切片的生物功能。结果亲和层析法纯化前小鼠腹腔积液蛋白质质量浓度平均为23．62mg／ml；纯化前、后蛋白总量分别为148．79和146．67mg，回收率为98．58％。腹腔积液中MAb质量浓度平均为5．21mg／ml；纯化的MAb纯度达95％以上。纯化后抗体免疫学活性均高于纯化前，提高了6．90～15．40倍。结论蛋白A-琼脂糖亲和层析法可快速、高效地从小鼠腹腔积液中纯化抗PGRPMAb，且抗体具有较高的纯度，免疫学活性明显提高，能够选择性的和小细胞肺癌细胞的PGRP结合。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.

关 键 词：色谱法 亲和 抗体 单克隆 纯化 

分 类 号：R734.2[医药卫生—肿瘤]