[Preparation and characterization of mono PEGylated recombinant human interferon omega]
| Autor: | Hong, Liu, Yong-Gang, Cheng, Hong-Chun, Pan, Bo, Xu, Li, Peng, Hong-Tao, Yang, Wei, Guo |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
Antigen-Antibody Reactions
Molecular Weight Drug Stability Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Interferon Type I Chromatography Gel Animals Electrophoresis Polyacrylamide Gel Rabbits Chromatography Ion Exchange Chromatography High Pressure Liquid Recombinant Proteins Polyethylene Glycols |
| Zdroj: | Yao xue xue bao = Acta pharmaceutica Sinica. 47(3) |
| ISSN: | 0513-4870 |
| Popis: | The amino group PEGylation of rhIFNomega with monomethoxy polyethylene glycol succinimidyl succinate (mPEG-SS, 20 000) was investigated, and the modified mixture was separated and purified by ion exchange chromatography and gel filtration chromatography. Under the optimized purification conditions, the average content ofmono PEG-rhIFNomega in the collect liquid reached 182 microg x mL(-1). The average purified yield of mono PEG-rhIFNomega exceed to 22%, and the purity of mono PEG-rhIFNomega was greater than 98% by SDS-PAGE and RP-HPLC. Relative molecular mass of mono PEG-rhIFNomega was 43 790 detected by MALDI-TOF MS. The apparent molecular mass measured by SDS-PAGE was about 60 810. The purified PEG-rhIFNomega has the characteristics of typical PEGylated protein. Activity reservation rate of mono PEG-rhIFNomega was 15.0%, while the antigenicity decreased by at least 64 folds. In addition, the acid stability, thermal stability and stability in serum and trypsin solution of mono PEG-rhIFNomega were markedly better than those of the rhIFNomega. The pharmacological properties of mono PEG-rhIFNomega were significantly improved. The prepared PEG-rhIFNomega might be developed to a novel safe and long-acting interferon. |
| Databáze: | OpenAIRE |
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