Evaluation of PEGylation reaction and purification of monoPEGylated recombinant human granulocyte colony stimulating factor
IR@NISCAIR: CSIR-NISCAIR, New Delhi - ONLINE PERIODICALS REPOSITORY (NOPR)
View Archive Info| Field | Value | |
| Creator |
Tiwari, Krishnanand
Kattavarapu, Krishna Shebannavar, Sunil N Pokalwar, Santosh. Mishra, Maheshwari K Chauhan, Ugam Kumari S |
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| Date |
2011-11-30T11:57:46Z
2011-11-30T11:57:46Z 2011-12 |
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| Identifier |
0975-1084 (Online); 0022-4456 (Print)
http://hdl.handle.net/123456789/13116 |
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| Description |
1049-1053
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. |
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| Language |
en_US
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| Publisher |
NISCAIR-CSIR, India
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| Rights |
<img src='http://nopr.niscair.res.in/image/cc-license-sml.png'> <a href='http://creativecommons.org/licenses/by-nc-nd/2.5/in' target='_blank'>CC Attribution-Noncommercial-No Derivative Works 2.5 India</a>
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| Source |
JSIR Vol.70(12) [December 2011]
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| Subject |
Bioassay
GCSF MPEG-ALD PEG GCSF, PEGylation SE-HPLC |
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| Title |
Evaluation of PEGylation reaction and purification of monoPEGylated recombinant human granulocyte colony stimulating factor
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| Type |
Article
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