CAJ | 학술논문

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%。
연구대장간균표체적중조인각질세포생장인자-Ⅰ형N단결실돌변체（△N23KGF-Ⅰ）적mPEG-마래선아알（mPEG-maleimide、mPEG-Mal）수식공예급수식위점화초보체외생물활성。수선수식득도취을이순화적△N23KGF-Ⅰ,연후대△N23KGF-Ⅰ작환원화비환원질량태도,학정기각태단,대△N23KGF-Ⅰ-m PEG-Mal-20K주환원질량태도,학정수식위점。최후,통과BAF-3세포대△N23KGF-Ⅰ국제표준품、△N23KGF-Ⅰ、△N23KGF-Ⅰ-m PEG-Mal-20K측활。결과표명,재뇨소배경하,mPEG-MAl피균일수식재△N23KGF-I적Cys40위。여국제활성표준품상비,△N23KGF-Ⅰ적활성보류료116%,△N23KGF-Ⅰ-m PEG-Mal-20K적활성보류료36%。
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%.