摘要
用化学合成法获得C末端氨基酸为半胱氨酸的Byetta类似物(ByC39),再采用分子量为40 kDa的马来酰亚胺活化单甲氧基聚乙二醇(mPEG-mal-40kDa)对ByC39的半胱氨酸巯基进行定点修饰,修饰产物通过阳离子交换层析和反向层析纯化获得。在已建立的定点修饰反应和分离纯化工艺基础上,通过改变反应条件中修饰pH值、修饰缓冲体系,以及修饰保护剂等因素来得到优化修饰条件:pH值为6.5的CH3COONa-CH3COOH缓冲液,保护剂为2.0mmol/L Lys和5.0 mmol/L EDTA,mPEG-mal-40 kDa/ByC39摩尔比3∶1,25℃反应4 h;经SDSPAGE和RP-HPLC检测,可知提高了修饰率以及增加了修饰产物的稳定性;最后加入TFA终止反应。
Obtained by chemical synthesis, the C-terminal amino acid is cysteine analogue Byetta （ByC39）. And then using a molecular weight of 40kDa maleimide-activated monomethoxy-polyethy- lene glycol （mPEG-mal-40kDa） on ByC39 sentinel cysteine ？？ thiol modification, modification prod- uct is purified by cation exchange chromatography and reverse chromatography to obtain. In the estab- lished point modification reaction and separation and purification process, based on the reaction condi- tions modified by changing pH, buffer system modification, and modification to optimize protection a- gents factors modified conditions： pH = 6.5 buffer of CH3 COONa-CH3 COOH, the protecting agent is2.0 mmol/L Lys and 5.0 mmot/L EDTA, mPEG-mal-40kDa/ByC39 reaction molar ratio 3： 1, 25℃ for 4 h. SDS-PAGE and RP-HPLC detection shows that the rate of modification and the addition modi- fied the stability of the product increase. TFA was added to terminate the reaction.
出处
《重庆理工大学学报（自然科学）》
CAS
2014年第2期45-48,共4页
Journal of Chongqing University of Technology：Natural Science
基金
国家重大新药创新"十一五"候选药物项目(2009ZX09103-6837)
关键词
促胰岛素分泌肽
聚乙二醇
修饰条件
glucogan-like peptide-1
polyethylene glycol
modified conditions
