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Site-specific PEGylation of Recombinant Human Non-glycosylated Erythropoietin and Characterization of the Mono-PEGylated Conjugate
Hao Su-Juan2; Wang Yin-Jue1,3; Kang Ai-Jun4; Liu Yong-Dong1; Li Xiu-Nan1; Shi Hong1; Ma Run-Yu2; Ma Guang-Hui1; Su Zhi-Guo1
关键词Erythropoietin Non-glycosylated Pegylation Mpeg-ald
刊名CHEMICAL JOURNAL OF CHINESE UNIVERSITIES-CHINESE
2010-11-10
31期:11页:2239-2245
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Chemistry, Multidisciplinary
研究领域[WOS]Chemistry
关键词[WOS]HAMSTER OVARY CELLS ; POLYETHYLENE-GLYCOL ; GROWTH-FACTOR ; IN-VITRO ; CARBOHYDRATE ; STABILITY ; HIRUDIN ; ANEMIA
英文摘要

Recombinant human erythropoietin (rhEpo) is a glycoprotein expressed in Chinese hamster ovary (CHO) cell. Carbohydrates play an important role in maintaining the protein′s stability and bioactivity. However, mammalian expressing system has low yields and high costs of production. In this article, a strategy of PEGylating E. coli expressed recombinant human non-glycosylated Epo (rh-ngEpo) by a 20000 site-specific monomethoxy polyethylene glycol propionaldehyde(mPEG-ALD) was investigated. The modification reaction was optimized and a high mono-modification yield of 55% was achieved. Ion exchange chromatography was then used to separate the monoPEGylated rh-ngEpo from the reaction mixture. The purity of the monoPEGylated rh-ngEpo was higher than 95% as indicated by HPSEC and RP-HPLC. The secondary and tertiary structures of rh-ngEpo were not changed by PEGylation. Rh-ngEpo was PEGylated mostly at the N-terminus by peptide mapping analysis. The in vitro bioactivity of the monoPEGylated rh-ngEpo decreased 30% compared with its unmodified counterpart while the thermal stability was greatly enhanced. The in vivo pharmacokinetic parameters were greatly enhanced. These results show that PEG could replace carbohydrates in enhancing the in vivo stability of nonglycosylated Epo. This research provides a direction for the development of new erythropoiesis-stimulating drugs.

语种中文
WOS记录号WOS:000285538100024
引用统计
被引频次:1[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/51921
专题实验动物科学部
北京大学医学部管理机构_医学部
作者单位1.Chinese Acad Sci, State Key Lab Biochem Engn, Inst Proc Engn, Beijing 100190, Peoples R China
2.Beijing Univ Chem Technol, Coll Life Sci & Technol, Beijing 100029, Peoples R China
3.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China
4.Peking Univ, Dept Lab Anim Sci, Hlth Sci Ctr, Beijing 100191, Peoples R China
推荐引用方式
GB/T 7714
Hao Su-Juan,Wang Yin-Jue,Kang Ai-Jun,et al. Site-specific PEGylation of Recombinant Human Non-glycosylated Erythropoietin and Characterization of the Mono-PEGylated Conjugate[J]. CHEMICAL JOURNAL OF CHINESE UNIVERSITIES-CHINESE,2010,31(11):2239-2245.
APA Hao Su-Juan.,Wang Yin-Jue.,Kang Ai-Jun.,Liu Yong-Dong.,Li Xiu-Nan.,...&Su Zhi-Guo.(2010).Site-specific PEGylation of Recombinant Human Non-glycosylated Erythropoietin and Characterization of the Mono-PEGylated Conjugate.CHEMICAL JOURNAL OF CHINESE UNIVERSITIES-CHINESE,31(11),2239-2245.
MLA Hao Su-Juan,et al."Site-specific PEGylation of Recombinant Human Non-glycosylated Erythropoietin and Characterization of the Mono-PEGylated Conjugate".CHEMICAL JOURNAL OF CHINESE UNIVERSITIES-CHINESE 31.11(2010):2239-2245.
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