Using continuous chromatography methodology to achieve high-productivity and high-purity enrichment of charge variants for analytical characterization

被引:12
作者
Bigelow, Elizabeth [1 ]
Song, Yuanli [1 ]
Chen, Jie [1 ]
Holstein, Melissa [1 ]
Huang, Yunping [2 ]
Duhamel, Lauren [1 ]
Stone, Kelly [1 ]
Furman, Ran [2 ]
Li, Zheng Jian [1 ]
Ghose, Sanchayita [1 ]
机构
[1] Bristol Myers Squibb, Biol Dev, 38 Jackson Rd, Devens, MA 01434 USA
[2] Bristol Myers Squibb, Biol Dev, 1 Squibb Dr, New Brunswick, NJ 08901 USA
关键词
Monoclonal Antibodies (mAbs); Charge Variant Enrichment; Multi-column Counter-current Solvent; Gradient Purification (MCSGP); Continuous Chromatography; Ion Exchange Chromatography; Analytical Characterization; CATION-EXCHANGE CHROMATOGRAPHY; PYROGLUTAMIC ACID; GLUTAMIC-ACID; HETEROGENEITY; ANTIBODIES; IGG1; SEPARATION; HPLC;
D O I
10.1016/j.chroma.2021.462008
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Charge variants of biological products, such as monoclonal antibodies (mAbs), often play an important role in stability and biological activity. Characterization of these charge variants is challenging, however, primarily due to the lack of both efficient and effective isolation methods. In this work, we present a novel use of an established, high productivity continuous chromatography method, known as multi column counter-current solvent gradient purification (MCSGP), to create an enriched product that can be better utilized for analytical characterization. We demonstrate the principle of this separation method and compare it to traditional batch HPLC (high performance liquid chromatography) or FPLC (fast protein liquid chromatography) methods, using the isolation of charge variants of different mAbs as a case study. In a majority of cases, we are able to show that the MCSGP method is able to provide enhanced purity and quantity of samples when compared to traditional fractionation methods, using the same separation conditions. In one such case, a sample prepared by MCSGP methodology achieved 95% purity in 10 hours of processing time, while those prepared by FPLC and HPLC achieved purities of 78% and 87% in 48 and 300 hours of processing time, respectively. We further evaluate charge variant enrichment strategies using both salt and pH gradients on cation exchange chromatography (CEX) and anion exchange chromatography (AEX) resins, to provide more effective separation and less sample processing following enrichment. As a result, we find that we are able to utilize different gradients to change the enrichment capabilities of certain charged species. Lastly, we summarize the identified mAb charge variants used in this work, and highlight benefits to analytical characterization of charge variants enriched with the continuous chromatography method. The method adds a new option for charge variant enrichment and facilitates analytical characterization of charge variants. (c) 2021 Elsevier B.V. All rights reserved.
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页数:9
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