聚乙二醇定点修饰haFGF突变体
취을이순정점수식haFGF돌변체
haFGF Mutant Site-directed Modified by PEG
  • 万方数据
    化学通报（网络版） 화학통보（망락판）
    Chemistry Online
    2004年 1期 ,共1页

    黄志锋%郑青*%苏志坚%吴晓萍%黄亚东%许华%赵文%李校坤%曲红艳%于平野

    황지봉%정청*%소지견%오효평%황아동%허화%조문%리교곤%곡홍염%우평야

    人酸性成纤维细胞生长因子突变体%聚乙二醇%定点修饰 人痠性成纖維細胞生長因子突變體%聚乙二醇%定點脩飾
    인산성성섬유세포생장인자돌변체%취을이순%정점수식
    Human acid fibroblast growth factor mutant%PEG%Site-directed modification
    将mPEG5000偶联到人酸性成纤维细胞生长因子(haFGF)突变体的第31位cys残基上,对haFGFSer98,132突变体进行定点修饰,修饰反应条件以马来酰亚胺-6-氨基己酸-mPEG5000酯和aFGFSer98,132摩尔比30/1、反应温度25℃、反应时间4h为佳.经反相高效液相色谱和SDS-PAGE凝胶电泳分析,修饰率达到30%以上,修饰后产物表观分子量为27kD,修饰产物的比活性保留55.53%.
    장mPEG5000우련도인산성성섬유세포생장인자(haFGF)돌변체적제31위cys잔기상,대haFGFSer98,132돌변체진행정점수식,수식반응조건이마래선아알-6-안기기산-mPEG5000지화aFGFSer98,132마이비30/1、반응온도25℃、반응시간4h위가.경반상고효액상색보화SDS-PAGE응효전영분석,수식솔체도30%이상,수식후산물표관분자량위27kD,수식산물적비활성보류55.53%.
    mPEG5000 was site-directed coupling to the 31st Cysteine residue of human acid fibroblast growth factor mutant, of which the 98th and the 132nd cysteine residue was replaced by Serine residue. The modified reaction was conducted at 25℃ for 4h. The optimal mol ratio of maleimide-6-aminocaproyl-mPEG5000 ester vs haFGFSer98,132 was 30/1. The modified products were analysed by reversed-phase HPLC and SDS-PAGE. The result indicated that the modification rate was up to 30% and the apparent molecular weight of the PEG-haFGF conjugate was 27kD. The final product PEG-haFGF was purified to homogeneity by Heparin affinity chromatography. The activity of the purified PEG-haFGF remained 55.53% of the activity of the haFGF mutant.
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