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Sodium caprylate wash during Protein A chromatography as an effective means for removing protease(s) responsible for target antibody fragmentation

Protein Expr Purif. 2021-05; 
Lixia Hu, Jiaqin Tang, Xudong Zhang, Yifeng Li
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Nucleic Acid Purification & Analysis … Louis, MO, USA). Precision Plus Protein Dual Color Standards was purchased from Bio-Rad Laboratories (Hercules, CA, USA). Precast NuPAGE 4–12% gradient Bis-Tris gels, 20X MES running buffer, 4X LDS sample buffer were purchased from GenScript (Nanjing, China)?… Get A Quote

摘要

For recombinant proteins produced in Chinese hamster ovary (CHO) cells, fragmentation is a common phenomenon that results in generation of product-related low-molecular-weight (LMW) species. Recently while purifying a bispecific antibody (bsAb), we observed that the target protein experienced cleavage at a couple of potential sites, leading to truncated products. Further studies suggest that the cleavage can likely be attributed to residual CHO cell protease activity. In order to maximally remove potential protease(s) that contribute fragmentation, we optimized Protein A chromatography by adding sodium caprylate (SC) to the wash buffer. Upon optimization, fragmentation of Protein A eluate happened to a much les... More

關鍵詞

Fragmentation, Host cell protein (HCP), Protease, Protein A chromatography, Sodium caprylate (SC)
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