摘要
目的对聚乙二醇(PEG)衍生物化学修饰重组人干扰素-α1b(rhIFN-α1b)的条件进行优化。方法以3种不同的第2代PEG衍生物对rhIFN-α1b进行修饰,SDS-PAGE分析PEG及修饰产物的表观相对分子质量。对反应pH值、温度、时间、修饰剂用量等修饰条件进行优化,并对修饰产物进行初步分离纯化和活性检测。结果PEG的电泳表观相对分子质量大于其理论值;PEG20000的修饰率(92.1%)高于PEG5000和PEG10000;修饰剂用量和pH值是主要影响因素,最佳pH为9.0,rhIFN-αlb与PEG20000的最佳摩尔比为1∶10,时间和温度影响甚微。单点修饰产物保留了14.4%的体外活性,多点修饰产物无活性。结论选择了合适的PEG衍生物并优化了修饰条件,为进一步深入研究奠定了基础。
Objective To optimize the condition for modification of recombinant human interferon-α1b (rhIFN-α1b) with polyethylene glycol (PEG). Methods Modify rhIFN-α1b with 3 kinds of PEG derivatives of the second generation, determine the apparent relative molecular masses of PEG and the modified rhIFN-α1b, and optimize the condition for modification, such as pH value, temperature, time and dosage of PEG.The modified rhIFN-α1b was primarily separated,purified and determined for activity. Results The apparent relative molecular mass of PEG was higher than that in theory. The modification rate of rhIFN-α1b with PEG20000(92.1% ) were significantly higher than that with PEG5000 or with PEG10000. The dosage of PEG and pH value showed significant influence, while time and temperature showed little influence on the modi- fication effect.The optimal pH value for modification was 9.0,and the optimal molar ratio of rhIFN-α1b to rhIFN-α1b to PEG20000 was 1 : 10.The mono-pegylated rhIFN-α1b reserved 14.4% of activity in vitro,while poly-pegylated rhIFN-α1b showed no activity. Conclusion The optimal PEG derivative for modification of rhIFN-α1b was screened,and the condition for modification was optimized, which laid a foundation of further study on modification of interferon with PEG.
出处
《中国生物制品学杂志》
CAS
CSCD
2008年第4期329-332,共4页
Chinese Journal of Biologicals
基金
国家863计划专题课题(2007AA02Z100)
上海市科委重点攻关计划(054319924)
关键词
聚乙二醇
重组人干扰素-α1b
化学修饰
优化
Polyethylene glycol
Recombinant human interferon-α1b
Chemical modification
Optimization