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Title: Evaluation of PEGylation reaction and purification of monoPEGylated recombinant human granulocyte colony stimulating factor
Authors: Tiwari, Krishnanand
Kattavarapu, Krishna
Shebannavar, Sunil N
Pokalwar, Santosh.
Mishra, Maheshwari K
Chauhan, Ugam Kumari S
Keywords: Bioassay;GCSF;MPEG-ALD;PEG GCSF,;PEGylation;SE-HPLC
Issue Date: Dec-2011
Publisher: NISCAIR-CSIR, India
Abstract: 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°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.
Page(s): 1049-1053
ISSN: 0975-1084 (Online); 0022-4456 (Print)
Appears in Collections:JSIR Vol.70(12) [December 2011]

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