Use and Optimization of a Dual-Flowrate Loading Strategy To Maximize Throughput in Protein-A Affinity Chromatography
详细信息 查看全文
- 作者：Sanchayita Ghose ; Deepak Nagrath ; Brian Hubbard ; Clayton Brooks ; and Steven M. Cramer
- 刊名：Biotechnology Progress
- 出版年：2004
- 出版时间：June 2004
- 年：2004
- 卷：20
- 期：3
- 页码：830 - 840
- 全文大小：265K
- 年卷期：v.20,no.3(June 2004)
- ISSN：1520-6033
文摘
The effect of an alternate strategy employing two different flowrates during loadingwas explored as a means of increasing system productivity in Protein-A chromatography. The effect of such a loading strategy was evaluated using a chromatographicmodel that was able to accurately predict experimental breakthrough curves for thisProtein-A system. A gradient-based optimization routine is carried out to establishthe optimal loading conditions (initial and final flowrates and switching time). Thetwo-step loading strategy (using a higher flowrate during the initial stages followedby a lower flowrate) was evaluated for an Fc-fusion protein and was found to result insignificant improvements in process throughput. In an extension of this optimizationroutine, dynamic loading capacity and productivity were simultaneously optimizedusing a weighted objective function, and this result was compared to that obtainedwith the single flowrate. Again, the dual-flowrate strategy was found to be superior.