Evaluation of PEGylation reaction and purification of monoPEGylated recombinant human granulocyte colony stimulating factor

被引：0

作者：

Tiwari, Krishnanand ^{[1
]}

Kattavarapu, Krishna ^{[1
]}

Shebannavar, Sunil N. ^{[1
]}

Pokalwar, Santosh ^{[1
]}

Mishra, Maheshwari K. ^{[1
]}

Chauhan, Ugam Kumari S. ^{[2
]}

机构：

[1] Gennova Biopharmaceut Ltd, MIDC, Pune 411057, Maharashtra, India

[2] Awdhesh Pratap Singh Univ, Dept Biotechnol, Rewa 486001, India

来源：

JOURNAL OF SCIENTIFIC & INDUSTRIAL RESEARCH | 2011年 / 70卷 / 12期

关键词：

Bioassay; GCSF; mPEG-ALD; PEG GCSF; PEGylation; SE-HPLC; AGGREGATION; PROTEIN; STABILITY; CSF; SEPARATION;

D O I：

暂无

中图分类号：

T [工业技术];

学科分类号：

08 ;

摘要：

In this study, incubation of methoxy polyethylene glycol propionaldehyde (mPEG-ALD) with recombinant human granulocyte colony stimulating factor (rh-GCSF) in presence of cyanoborohydride yielded various species of polyethylene glycol (PEG) conjugated GCSF as detected by size exclusion high performance liquid chromatography (SE-HPLC). These reactions were carried out at 1:5 molar ratio of rh-GCSF to PEG at 20-25 degrees C. At 1 h of reaction, mainly monoPEGylated GCSF (62.4%) was formed. At 2 h of reaction, multi PEGylated GCSF (conc. 9%) was detected, where monoPEGylated GCSF constituted the most (73.2%). At 3 h of reaction, monoPEGylated (76.2%) and multi PEGylated GCSF (11.3%) were formed. Further, a process involving ion exchange and gel filtration chromatography were used to obtain pure monoPEGylated GCSF. Purified monoPEGylated GCSF was comparable to standard PEGylated rh-GCSF on NFS-60 cell line, suggesting retained biological activity of monoPEGylated GCSF. Further, the procedure is warranted for purification of other monoPEGylated proteins for therapeutic purpose.

引用

页码：1049 / 1053

页数：5

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