Liu H.,Southwest University |
Cheng Y.-G.,Southwest Pharmaceuticals Ltd Company |
Pan H.-C.,Southwest University |
Xu B.,Southwest Pharmaceuticals Ltd Company |
And 3 more authors.
Yaoxue Xuebao | Year: 2012
The amino group PEGylation of rhIFNω 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 of mono PEG-rhIFNω in the collect liquid reached 182 μg·mL-1. The average purified yield of mono PEG-rhIFNω exceed to 22%, and the purity of mono PEG-rhIFNω was greater than 98% by SDS-PAGE and RP-HPLC. Relative molecular mass of mono PEG-rhIFNω was 43 790 detected by MALDI-TOF MS. The apparent molecular mass measured by SDS-PAGE was about 60 810. The purified PEG-rhIFNω has the characteristics of typical PEGylated protein. Activity reservation rate of mono PEG-rhIFNω 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-rhIFNω were markedly better than those of the rhIFNω. The pharmacological properties of mono PEG-rhIFNω were significantly improved. The prepared PEG-rhIFNω might be developed to a novel safe and long-acting interferon. Source