摘要
研究L-门冬酰胺酶的单甲氧基聚乙二醇醛化学修饰及纯化方法。以SDS-PAGE分析产物的单修饰产率,对反应的摩尔比,反应的pH值和反应时间和温度修饰条件进行优化,初步确定最佳修饰条件为L-ASP与mPEG、ALD反应摩尔比是1∶10,反应pH值为5.0,在4℃下反应32 h。离子交换和分子筛联用对产物进行分离纯化,修饰产物活性残留达36%。修饰产物结构没有明显变化,活性得到很好的保留。
L-asparaginase (L-ASP) modified with succinimidehyde monomethoxy polyethylene glycol (mPEG-ALD) and its purification was studied. The modification rate of mono-PEGylated enzyme by SDS-PAGE was analyzed and the modified conditions, such as molecular ratio, pH value, reaction tem- perature and time were optimized. The optimized reaction conditions were found to be phosphate buffer pHS. 0,moleeuar ratio of mPEG-ALD to L-ASP 10 : 1, reaction time 32h and reaction temperature 4℃. The PEGylated enzyme was purified by anion-exchange and gel filtration and its activity retained 360%. The modified PEG-L-ASP with noob change in strueture and its activity has been well reserved.
出处
《药物生物技术》
CAS
CSCD
2009年第3期212-216,共5页
Pharmaceutical Biotechnology
基金
校内药物传递系统技术平台项目(35210514)
关键词
L-门冬酰胺酶
聚乙二醇醛
化学修饰
分离纯化
L - asparaginase, mPEG-ALD, Chemical modification, Isolation and purification