期刊
PROCESS BIOCHEMISTRY
卷 55, 期 -, 页码 162-171出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.procbio.2017.01.024
关键词
Recombinant hirudin; PEGylation; Ion exchange chromatography; In situ PEGylation; Solid-phase PEGylation; On-column PEGylation
资金
- National Natural Science Foundation of China [81072590, 21606038]
- Jiangsu Province State Key Laboratory Cultivating Base for Long-acting Bio-medical Research of Jiangsu Hansoh Pharmaceutical Group CO., LTD
In this study, an integrated process was developed for successive solid-phase PEGylation of recombinant hirudin variant-2 (HV2) and separation of PEGylated HV2 species on an anion exchange chromatography column (so-called in situ PEGylation). The effects of different PEG sizes, ion exchange resins and reaction conditions on in situ PEGylation were investigated. The results showed that in situ PEGylation efficiently integrates the reaction, separation and purification into a single-unit operation using the same column. In situ PEGylation could improve the selectivity of PEGylation reactions by significantly reducing the formation of multi-PEG-HV2. The pore sizes and internal surface structures of different resins had a significant impact on the yield of mono-PEG-HV2. In contrast to liquid-phase PEGylation, the yield of mono-PEG-HV2 decreased as PEG size increased during the in situ PEGylation process, indicating that in situ PEGylation is a pore diffusion-controlled process. The in vitro and in vivo anticoagulant activities of mono-PEG-HV2 derived from in situ PEGylation were higher than those from liquid-phase PEGylation, indicating that in situ PEGylation could enhance the bioactivity retention of mono-PEG-HV2. The results of this study demonstrated that in situ PEGylation can be used as an effective approach for the development of PEGylated protein drugs. (C) 2017 Elsevier Ltd. All rights reserved.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据