Site-specific PEGylation of Human Growth Hormone by Mutated Sortase A  

作　　者：SHI Hui SHI Qingyang James T. OSWALD GAO Ying LI Leijiao LI Yunhui 

机构地区：[1]School of Chemistry & Environmental Engineering, Changchun University of Science & Technology, Changchun 130022, P R. China [2]Key Laboratory of Regenerative Biology of the Chinese Academy of Sciences Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, P. R. China [3]University of Chinese Academy of Sciences, Beijing P. R. China,100049 [4]Centre for Reproductive Medicine, Centre for Prenatal Diagnosis, the First Hospital of Jilin Universi(y. Changchun 130021, P. R. China [5]School of Nanoteehnology & Chemical Engineering, University of Waterloo, Waterloo ON, Canada

出　　处：《Chemical Research in Chinese Universities》2018年第3期428-433,共6页高等学校化学研究（英文版）

基　　金：Supported by the Projects of Science ＆ Technology of Jilin Province, China（No.20150204030YY）, the Project of the Research and Development of Industrial Technology of Jilin Province, China（No.2015Y058） and the National Natural Science Foundation of China（No. 51402286 ）.

摘　　要：Human growth hormone（hGH）, a classic therapeutic protein, which promotes growth and wound healing, is released from the pituitary gland. As a protein drug, its short half-life is its main barrier to therapeutic efficacy. Various strategies have been designed to prolong its serum half-life, the most conunon of which is the conjugation with polyethylene glycol（PEG）, as this has been shown to significantly extend protein＇s serum half-life. However, PEGylation often results in random conjugation, which can lead to impaired protein function and hinder purification, characterization and evaluation of the PEGylated protein. Therefore, site specific PEGylation is a promising direction for PEG-protein conjugation. Here we took advantages of the mutated sortase A（7M） enzyme, which can enzymati- cally ligate the universal a-amino acids to a C-terminal tagged protein. This then allows specific modification of the C-terminal of hGH with PEG. This site-specific bound PEG-hGH has similar efficacy, receptor binding and cell proliferation as wild-type hGH; however, pharmacokinetic analysis demonstrates that its serum half-life is almost 24 times that of wild-type hGH. Herein, we provided a promising advancement in the development of site specific PEGylated therapeutic proteins.Human growth hormone（hGH）, a classic therapeutic protein, which promotes growth and wound healing, is released from the pituitary gland. As a protein drug, its short half-life is its main barrier to therapeutic efficacy. Various strategies have been designed to prolong its serum half-life, the most conunon of which is the conjugation with polyethylene glycol（PEG）, as this has been shown to significantly extend protein＇s serum half-life. However, PEGylation often results in random conjugation, which can lead to impaired protein function and hinder purification, characterization and evaluation of the PEGylated protein. Therefore, site specific PEGylation is a promising direction for PEG-protein conjugation. Here we took advantages of the mutated sortase A（7M） enzyme, which can enzymati- cally ligate the universal a-amino acids to a C-terminal tagged protein. This then allows specific modification of the C-terminal of hGH with PEG. This site-specific bound PEG-hGH has similar efficacy, receptor binding and cell proliferation as wild-type hGH; however, pharmacokinetic analysis demonstrates that its serum half-life is almost 24 times that of wild-type hGH. Herein, we provided a promising advancement in the development of site specific PEGylated therapeutic proteins.

关 键 词：Human growth hormone PEGYLATION Site specific modification 

分 类 号：Q57[生物学—生物化学] Q51