重组抗人CD25嵌合单克隆抗体的表达及纯化  

Expression and Purification of Recombinant Anti-human CD25 Chimeric Monoclonal Antibody

作　　者：王进[1,2] 张敏 孙广瑞 刘莹 熊思东[2] 

机构地区：[1]上海新生源生物医药有限公司,上海201203 [2]复旦大学免疫生物学研究所,上海200032

出　　处：《中国医药工业杂志》2009年第11期818-821,共4页Chinese Journal of Pharmaceuticals

基　　金：上海市科委重点科技攻关项目(074319108)

摘　　要：采用NS0工程细胞表达重组抗人CD25嵌合单克隆抗体,在5L细胞反应器进行批次培养,细胞悬液中目标单抗的平均表达量为108mg/L,每批细胞悬液总体积为4L。细胞悬液离心后上清液经rProtein A Sepharose FF和SP Sepharose FF两步色谱纯化,得纯度大于95%的单抗,回收率约33%。纯化产物经Western blotting验证为目标单抗,具有免疫原性。N-端15个氨基酸序列测定结果与理论序列相符。The engineered NS0 myeloma cell line was used to express recombinant anti-human CD25 chimeric monoclonal antibody. The cell cultivation and expression process was investigated in a 5 L cell reactor. The average yield of target product in cell suspensions was 108 mg/L, and about 4 L cell suspensions were harvested from each batch. The target product with purity over 95 % and recovery rate around 33 % was purified from harvested supernatant by a two-step purification process using rProtein A Sepharose FF and SP Sepharose FF chromatography. Western blotting proved that the expressed and purified protein was identical to the target product and retained the immunogenicity of the target product. Analysis result showed that fifteen amino acid residues of the N-terminal were identical to the theoretical sequence.

关 键 词：抗人CD25 嵌合单克隆抗体 细胞表达 纯化 

分 类 号：Q511[生物学—生物化学]