[Preparation and characterization of mono PEGylated recombinant human interferon omega].
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.
School of Pharmaceutical Sciences, Southwest University, Chongqing 400716, China.
Hong
H
Liu
Yong-Gang
YG
Cheng
Hong-Chun
HC
Pan
Bo
B
Xu
Li
L
Peng
Hong-Tao
HT
Yang
Wei
W
Guo
D000818:Animals; D000937:Antigen-Antibody Reactions; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004355:Drug Stability; D004591:Electrophoresis, Polyacrylamide Gel; D007370:Interferon Type I / Q000737:chemistry* / Q000276:immunology; D008970:Molecular Weight; D011092:Polyethylene Glycols / Q000737:chemistry*; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D007370:Interferon Type I; D011994:Recombinant Proteins; C063743:interferon omega 1; D011092:Polyethylene Glycols; C028210:monomethoxypolyethylene glycol