重组抗人CD25嵌合单克隆抗体的表达及纯化

采用NSO工程细胞表达重组抗人CD25嵌合单克隆抗体,在5 L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108 mg/L,每批细胞悬液总体积为4 L.细胞悬液离心后上清液经rProtein A Sepharose FF和SPSepharose 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.

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

The engineered NS0 myeloma cell line was used to express recombinant anti-human CD25 chimeric monocional 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.