高效亲和色谱法检测细胞培养上清中的抗体融合蛋白质

Determination of antibody fusion protein in cell culture supernatant by high performance affinity chromatography

目的建立高效亲和色谱检测抗体融合蛋白质的方法，用于抗体及抗体融合蛋白质生产的过程控制。方法正压匀浆法装填rProtein A Sepharose，用不同pH值缓冲液分步洗脱。结果在25～1000μg／mL浓度范围内，重组人肿瘤坏死因子受体一抗体融合蛋白（TNFR—Fc）浓度和峰面积显著相关，r=0.9996；回收率范围为91％～99％，对表达TNFR—Fc的CHO工程细胞的不同培养阶段的3份上清分别进行3次重复测定，RSD在2％以内。结论该方法适用于抗体及抗体融合蛋白质生产的过程控制。

Purpose The method for determination of antibody fusion protein was developed for process control of antibody or antibody fusion protein production. Methods The column used for high performance affinity chromatography was packed by positive pressure with rProtein A Sepharose. The samples were step eluted with buffer with different pH. Results The concentration of TNFR-Fc was correlated well with the peak area, r=0. 999 6. The relative standard deviation of determination to three culture supernants from genetic CHO cell expressing TNFR-Fc was less than 2%. Conclusion The established high performance affinity chromatography is suitable for process control of antibody fusion protein production.