3种不同Protein A亲和层析填料纯化抗CD52单克隆抗体的比较

目的 比较3种Protein A亲和层析填料MabSelect SuRe、Protein A Diamond及UniMab 50纯化抗CD52单克隆抗体的载量及纯化效果.方法 将浓度为1.24 mg/mL的抗CD52单克隆抗体分别流经3种Protein A亲和层析填料,确保保留时间一致,检测流穿液的抗体浓度,以流穿液中...

Purification of anti-CD52 monoclonal antibody with three media for protein A affinity chromatography

Objective To compare the capacity and purification effect of three media,MabSelect SuRe,Protein A Diamond and UniMab 50,in purification of anti-CD52 monoclonal antibody(McAb).Methods The anti-CD52 McAb at a concentration of 1.24 mg/mL was purified by affinity chromatography with the three media respectively,and the same retention times were ensured.The antibody concentration of the flowing through liquid was measured.The maximum dynamic loading of the medium was calculated with the sample loading when the antibody concentration in flowing through liquid reached 10% of the sample concentration.Three batches of culture supernatant of McAb against CD52 were purified with the three media at 80% of the maximum loading,and the elution volume,antibody recovery as well as the concentration and purity of antibody,charge heterogeneity,residual host protein content,residual host DNA content and protein A abscission in the eluate were compared.Results MabSelect SuRe was similar to Protein A Diamond in terms of loading capacity,residual host protein content and protein A abscission.The three media showed the same performance in terms of purity,charge heterogeneity and residual host DNA content.However,unimab 50 showed the best performance in antibody recovery,elution volume and antibody concentration.Conclusion The three media showed either advantages and disadvantages in the purification of anti-CD52 McAb by affinity chromatography,all of which met the requirement for purification in this step and might be selected according to the practical process conditions and requirements for quality control.