双柱连续流层析亲和分离抗体的过程设计与应用

Process design and application of twin-column continuous chromatography for antibody affinity separation

针对单抗生产下游过程的瓶颈-蛋白A亲和分离,以提高产率和降低成本为目标,探讨了双柱连续流层析的新方法。从过程设计的关键因素出发,比较和分析了保留时间和蛋白浓度的影响,优化了分离条件,并应用于细胞培养液中单抗的分离。以两种蛋白A亲和介质MabSelect SuRe和UniMab为典型介质,考察了不同保留时间和IgG浓度下的穿透曲线,比较了保留时间和蛋白浓度对产率、介质利用率和缓冲液消耗的影响。结果表明,与常规的批次层析相比,双柱连续流层析拥有更优越的分离性能,低保留时间条件下的产率和介质利用率显著提高,缓冲液消耗减少,特别适合于高流速操作。对于从细胞培养液中分离单抗,双柱连续流层析的分离性能与常规批次层析相当,单抗纯度在97%以上、收率87%以上、聚集体含量低于2%、宿主细胞蛋白含量低于300ppm,优势是产率提高,介质利用率从20%～30%提高到70%～80%,,缓冲液消耗下降为常规批次层析过程的1/5～1/4,显示出良好的应用前景。

In order to improve process throughput and reduce operation cost of Protein A affinity chromatography,a twin-column continuous chromatography method was studied for monoclonal antibody(mAb)separation.Effects of residence time in the column and loading protein concentration were investigated,and the separation conditions were optimized for mAb capture from cell culture broth.Two typical Protein A affinity resin(MabSelect SuRe and UniMab)were used and the breakthrough curves were measured under different residence times and IgG concentrations,and their effects on the productivity,resin capacity utilization and buffer consumption were evaluated.The results indicate that the twin-column continuous chromatography method has higher productivity and capacity utilization,and lower buffer consumption when comparing to traditional batch chromatography at low residence times.The separation performance of continuous chromatography for mAb capture from cell culture broth was similar to batch chromatography with purity >97%,yield >87%,aggregates<2%and host cell proteins<300 ppm.However,the capacity utilization increased from 20%~30%to 70%~80%,and the buffer consumption reduced to 1/5~1/4 of traditional batch processes.The results demonstrate that the twin-column continuous chromatography method shows great potential for the mAb production.