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Journal of Scientific and Industrial Research (JSIR) >
JSIR Vol.70 [2011] >
JSIR Vol.70(12) [December 2011] >
| 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 |
| CC License: |  CC Attribution-Noncommercial-No Derivative Works 2.5 India |
| ISSN: | 0975-1084 (Online); 0022-4456 (Print) |
| Source: | JSIR Vol.70(12) [December 2011]
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