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犬干扰素-γ的稀释复性及活力测定
Renaturation and activity determination of recombinant canine interferon-γ inclusion body by dilution
【摘要】 文章探讨了不同折叠促进剂、加样方式、温度、pH在稀释法复性条件下对重组犬IFN-γ体外折叠的复性影响。结果表明,0.5mol·L-1精氨酸、0.5mol·L-1盐酸胍、2.0mol·L-1尿素均能有效抑制复性过程中蛋白质的聚集,其中精氨酸的效果最好。在4℃、pH8.0、精氨酸浓度0.5mol·L-1,蛋白浓度0.15mg·mL-1及脉冲加样操作方式下,复性后重组犬IFN-γ的活性为1.35×104U·mL-1,比活为3.74×106U·mg-1。
【Abstract】 Effects of folding enhancer, ways of adding sample, temperature and pH were investigated in the renatutation process of recombinant canine interferon-gamma inclusion body by dilution. The results showed that 0.5 mol·L-1 L-arginine, 0.5 mol·L-1 guanidine-HCl and 2.0 mol·L-1 urea could suppress the aggregation of protein during the renaturation process effectively, and 0.5 mol·L-1 L-arginine was the best of them. The activity and the specific bioactivity were up to 1.35×104 U·mL-1 and 3.74×106 U·mg-1, respectively under 4 ℃, pH 8.0, 0.5 mol·L-1 L-arginine, 0.15 mg·mL-1 proein and pulse renturation operation mode.
【Key words】 recombinant canine interferon-γ; renaturaton by dilution; aggregation suppressors; activity; specific bioactivity; inclusion body;
- 【文献出处】 东北农业大学学报 ,Journal of Northeast Agricultural University , 编辑部邮箱 ,2008年09期
- 【分类号】S852.4
- 【被引频次】14
- 【下载频次】251