Cathepsin L Causes Proteolytic Cleavage of Chinese-Hamster-Ovary Cell Expressed Proteins During Processing and Storage: Identification, Characterization, and Mitigation

Luo Haibin; Tie Liu; Cao MingyanHunter Alan K.Pabst Timothy M.Du JialiField RaymondLi YulingWang William K.

首页> 外文期刊>Biotechnology Progress >Cathepsin L Causes Proteolytic Cleavage of Chinese-Hamster-Ovary Cell Expressed Proteins During Processing and Storage: Identification, Characterization, and Mitigation

【24h】

Cathepsin L Causes Proteolytic Cleavage of Chinese-Hamster-Ovary Cell Expressed Proteins During Processing and Storage: Identification, Characterization, and Mitigation

机译：Cathepsin L Causes Proteolytic Cleavage of Chinese-Hamster-Ovary Cell Expressed Proteins During Processing and Storage: Identification, Characterization, and Mitigation

获取原文

获取原文并翻译 | 示例

掌桥外文数据库（机构版） >>

开具论文收录证明 >>

文献代查 >>

页面导航

摘要
著录项
相关主题

摘要

A stochastic approach of copurification of the protease Cathepsin L that results in product fragmentation during purification processing and storage is presented. Cathepsin L was identified using mass spectroscopy, characterization of proteolytic activity, and comparison with fragmentation patterns observed using recombinant Cathepsin L. Cathepsin L existed in Chinese hamster ovary cell culture fluids obtained from cell lines expressing different products and cleaved a variety of recombinant proteins including monoclonal antibodies, antibody fragments, bispecific antibodies, and fusion proteins. Therefore, characterization its chromatographic behavior is essential to ensure robust manufacturing and sufficient shelf life. The chromatographic behaviors of Cathepsin L using a variety of techniques including affinity, cation exchange, anion exchange, and mixed mode chromatography were systematically evaluated. Our data demonstrates that copurification of Cathepsin L on nonaffinity modalities is principally because of similar retention on the stationary phase and not through interactions with product. Lastly, Cathespin L exhibits a broad elution profile in cation exchange chromatography (CEX) likely because of its different forms. Affinity purification is free of fragmentation issue, making affinity capture the best mitigation of Cathepsin L. When affinity purification is not feasible, a high pH wash on CEX can effectively remove Cathepsin L but resulted in significant product loss, while anion exchange chromatography operated in flow-through mode does not efficiently remove Cathepsin L. Mixed mode chromatography, using Capto (TM) adhere in this example, provides robust clearance over wide process parameter range (pH 7.7 +/- 0.3 and 100 +/- 50 mM NaCl), making it an ideal technique to clear Cathepsin L. (c) 2018 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2732, 2019

著录项

来源
《Biotechnology Progress》 |2019年第1期|2732-1-2732-12|共12页
作者
Luo Haibin; Tie Liu; Cao MingyanHunter Alan K.Pabst Timothy M.Du JialiField RaymondLi YulingWang William K.;
展开▼
作者单位

Medimmune LLC, Purificat Proc Sci, Gaithersburg, MD 20878 USA;

Medimmune LLC, Analyt Sci, Gaithersburg, MD 20878 USA;

Medimmune LLC, Formulat Sci, Gaithersburg, MD 20878 USAMedimmune LLC, Cell Culture & Fermentat Sci, Cambridge CB21 6GH, England;

展开▼
收录信息
原文格式 PDF
正文语种英语
中图分类生物科学;
关键词
fragmentation; host cell proteins; cysteine protease; Cathepsin L; copurification;

Cathepsin L Causes Proteolytic Cleavage of Chinese-Hamster-Ovary Cell Expressed Proteins During Processing and Storage: Identification, Characterization, and Mitigation

摘要

著录项

相关主题

期刊订阅