Solid Phase Pegylation of Hemoglobin
A solid phase conjugation process was developed for attachment of polyethylene glycol to hemoglobin molecule. Bovine hemoglobin was loaded onto an ion exchange chromatography column and adsorbed by the solid medium. Succinimidyl carbonate mPEG was introduced in the mobile phase after the adsorption....
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Published in | Artificial cells, blood substitutes, and immobilization biotechnology Vol. 37; no. 4; pp. 147 - 155 |
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Main Authors | , , , , , |
Format | Journal Article |
Language | English |
Published |
England
Informa UK Ltd
01.01.2009
Taylor & Francis |
Subjects | |
Online Access | Get full text |
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Summary: | A solid phase conjugation process was developed for attachment of polyethylene glycol to hemoglobin molecule. Bovine hemoglobin was loaded onto an ion exchange chromatography column and adsorbed by the solid medium. Succinimidyl carbonate mPEG was introduced in the mobile phase after the adsorption. Pegylation took place between the hemoglobin on the solid phase, and the pegylation reagent in the liquid phase. A further elution was carried out to separate the pegylated and the unpegylated protein. Analysis by HPSEC, SDS-PAGE, and MALLS demonstrated that the fractions eluted from the solid phase contained well-defined components. Pegylated hemoglobin with one PEG chain was obtained with the yield of 75%, in comparison to the yield of 30% in the liquid phase pegylation. The P50 values of the mono-pegylated hemoglobin, prepared with SC-mPEG 5kDa, 10kDa and 20kDa, were 19.97, 20.23 and 20.54 mmHg, which were much closer to the value of red blood cells than that of pegylated hemoglobin prepared with the conventional method. |
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Bibliography: | ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23 ObjectType-Article-1 ObjectType-Feature-2 |
ISSN: | 1073-1199 1532-4184 |
DOI: | 10.1080/10731190903041188 |