摘要
研究大肠杆菌表达的重组人角质细胞生长因子-Ⅰ型N端缺失突变体(△N23KGF-Ⅰ)的mPEG-马来酰亚胺(mPEG-maleimide、mPEG-Mal)修饰工艺及修饰位点和初步体外生物活性。首先修饰得到聚乙二醇化的△N23KGF-Ⅰ,然后对△N23KGF-Ⅰ作还原和非还原质量肽图,确定其各肽段,对△N23KGF-Ⅰ-m PEG-Mal-20K做还原质量肽图,确定修饰位点。最后,通过BAF-3细胞对△N23KGF-Ⅰ国际标准品、△N23KGF-Ⅰ、△N23KGF-Ⅰ-m PEG-Mal-20K测活。结果表明,在尿素环境下,mPEG-MAl被均一修饰在△N23KGF-I的Cys40位。与国际活性标准品相比,△N23KGF-Ⅰ的活性保留了116%,△N23KGF-Ⅰ-m PEG-Mal-20K的活性保留了36%。
The research studied the PEGylation of mPEG-maleimide （mPEG-Mal） of recombinant ke- ratinocyte growth factor-I mutant deleted on N-terminus （ △N23KGF-I） expressed in E. coli, and i- dentified the modified site and preliminary biological activity in vitro. Firstly, get the Pegylated AN23KGF-I; Then, identify all the segments of the peptide by the reduced and non-reduced peptide mass mapping （PMM）. Next, identify the modified site of △23KGF-I-mPEG-Mal-20K by the reduced PMM. Lastly, measure the activity of △N23KGF- I international standard product, △N23KGF-I, △N23KGF-I-mPEG-Mal-20K with BAF-3 cell line. And mPEG-Mal-20K is modified uniformly on Cys40 when urea exists. In contrast to △N23KGF-I international standard product, the activity of △N23KGF-I keeps about 116%, and the △N23KGF-I-m PEG-Mal-20K keeps about 36%.
出处
《重庆理工大学学报（自然科学）》
CAS
2012年第4期40-44,共5页
Journal of Chongqing University of Technology：Natural Science
基金
国家科技重大专项资助项目(2012ZX09103-301-052)
重庆市自然科学基金资助项目(CSTC2007BB5412)
