一种利用HCIC快速纯化抗rhTF_(243)单克隆抗体方法的研究  

Study on fast purification of monoclonal antibody against rhTF_(243) protein with the technique of HCIC

作　　者：赵峰梅[1] 齐延红[1] 何永吉[1] 潘薇薇[1] 卢晓霞[1] 粱雅丽 赵邑[1] 

机构地区：[1]山西省生物研究所,太原030006

出　　处：《中国免疫学杂志》2010年第6期548-551,共4页Chinese Journal of Immunology

基　　金：山西省国际科技合作资助项目(No.20081041)

摘　　要：目的:建立从小鼠腹水中快速纯化抗人重组组织因子243(recombine human tissue factor243,rhTF243)单克隆抗体(monoclonal antibody,mAb)的方法。方法:含抗rhTF243单克隆抗体的腹水经离心、过滤及缓冲溶液变换等预处理后,先经疏水电荷诱导层析纯化处理,去除大部分杂蛋白,再经Protein A-Sepharose CL-4B亲和层析柱进一步纯化。结果:经两步纯化后获得抗rhTF243单克隆抗体,其生物学活性高、纯度大于95%,抗体效价保持良好,抗体总回收率达73%,且具有明显的抗促凝作用。结论:建立的单克隆抗体纯化方法简便、高效,所得mAb的纯度高、生物学活性好。Objective：To develop a two-step method for purification of monoclonal antibody rhTF243 protein from mouse ascites by using hydrophobic charge induction chromatography（HCIC） and affinity chromatography with protein A sepharose CL-4B.Methods：The ascites was first purified by HCIC after centrifugation and filtration.Then the fraction containing the protein of interest was directly purified by affinity chromatography with protein A sepharose CL-4B.Results：The purity of the obtained monoclonal antibody was up to 97% with recovery of 73% and of high activity.Conclusion：The method for purification of monoclonal antibody is developed using HCIC and Protein A affinity chromatography and the obtained antibodies are of high purity and activity.

关 键 词：MEP HyperCel疏水层析 PROTEIN A-Sepharose CL-4B亲和层析 单克隆抗体 腹水 纯化 

分 类 号：R392.11[医药卫生—免疫学]