期刊
BIOCHEMICAL ENGINEERING JOURNAL
卷 131, 期 -, 页码 47-57出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.bej.2017.12.001
关键词
High-throughput screening; Mixed-mode chromatography; Microtiter filter plate; Human serum albumin; Protein separation
资金
- International Science & Technology Cooperation Program of China
- National Natural Science Foundation of China
High-throughput approaches become increasingly important on process development for protein separation. In this work, four mixed-mode resins (Capto adhere, Capto MMC, MX-Trp-650 M and Nuvia cPrime) were investigated for human serum albumin (HSA) separation and compared with two ion-exchangers (Q Sepharose FF and SP Sepharose FF). High-throughput method based on microtiter filter plate was used to determine adsorption properties and optimize separation conditions. For resins with anion-exchange groups alkaline condition would be favorable for HSA adsorption, while acidic condition for resins with cation-exchange groups. Mixed-mode resins showed obviously salt-tolerant properties. Salt addition had little effect on Nuvia cPrime, and could promote HSA adsorption for Capto MMC and MX-Trp-650 M. The elution conditions were also optimized and high recovery could be obtained. In addition, the optimized conditions obtained by high-throughput approaches were verified by lab-scale column chromatography. It was found that Capto MMC and MX-Trp-650 M would be more suitable for HSA separation and DBC10% could reach more than 40 mg/ml under 200 mM NaCl addition. Finally, rHSA separation from P. pastoris culture broth was tested with MX-Trp-650 M, and high purity and recovery could be obtained. This study demonstrated that high-throughput screening method is a powerful tool to optimize the chromatographic separation. Mixed-mode cation-exchange resins showed an efficient practicability for HSA separation with high capacity, suitable salt-tolerance and high elution recovery. (C) 2017 Elsevier B.V. All rights reserved.
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