Process Design And Antibody Separation With Twin-column Continuous Chromatography

Monoclonal antibodies(mAbs)are one of the most important biopharmaceuticals.The current antibody purification platform based on Protein A affinity chromatography cannot meet the huge market demand as the limited productivity.Therefore,it is necessary to develop new sepration methods for mAb manufacuring.Continuous chromatography is new separation technology as one important part of continuous biomanufacturing,but the research on the process factors is limited.The twin-column continuous chromatography was focused for antibody capture in this thesis.Based on the key operation factors,the process performance of traditional batch chromatography and continuous chromatography were investigated.Several Protein A affinity resins were tested and compared.The results would giude the process development and applications of continuous chromatography.With the typical Protein A resin MabSelect SuRe and two new Protein resins(Resin A and Resin B)as the model,the effects of residence time and protein concentration on the process of twin-column continuous chromatography were investigated.The results indicated that with the increase of residence time the process productivity of three resins increased first and then decreased,and the capacity utilization increased gradually.The capacity utilization of twin-columns continuous chromatography(more than 70%)was much higher than that of batch chromatography(20-50%),and the buffer consumption was reduced by more than 50%.As the protein concentration increased,the productivity increased significantly,the capacity utilization and buffer consumption remained constant.Among the three resins tested,Resin B has the highest loading capacity and the best separation performance.The productivity reached 43.0 g/(L·h)at the residence time of 1 min.The separation processes of mAb from cell culture broth were studied with twin-column continuous chromatography.The results indicated that mAb quality were similar to traditional atch chromatography process,The mAb purity was above 98%,with the aggregates content between 1%and 2.5%and HCP content about 200?600 ppm.Among the three resins tested,Resin B has the best separation perfonnances with the recovery of 96%,the productivity of 37.5 g/(L h).Compared with batch process,the buffer consumption was reduced 63%.The effect of column height were evaluated,and it was found that the shorter column has better performance.Finally,the AutoMab loading control was tested.The results showed that the process was stable.The amount of protein eluted in each cycle was similar,and the difference in concentration was less than 3%.The mAb purity reached 98%.The results demonstrated that twin-column continuous chromatography is feasible for the mAb production.Aiming at antibody separation with twin-column continuous chromatography,the effects of operationt conditions on the process performance were investigated.The operating parameters were optimized and applied successfully to mAb capture from cell culture broth.The process design method developed in this thesis would be useful for the process development of continuous chromatography.