| 犬干扰素-γ的稀释复性及活力测定 |
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| 引用本文： | 陈忠广,张桂红,夏春丽,李宝贤,李一经.犬干扰素-γ的稀释复性及活力测定[J].东北农业大学学报,2008,39(9). |
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| 作者姓名： | 陈忠广 张桂红 夏春丽 李宝贤 李一经 |
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| 作者单位： | 东北农业大学动物医学学院,哈尔滨,150030 |
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| 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。
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| 关 键 词： | 重组犬IFN-γ 稀释复性 折叠促进剂 活性 比活 包涵体 |
Renaturation and activity determination of recombinant canine interferon-γ inclusion body by dilution |
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| CHEN Zhongguang,ZHANG Guihong,XIA Chunli,LI Baoxian,LI Yijing.Renaturation and activity determination of recombinant canine interferon-γ inclusion body by dilution[J].Journal of Northeast Agricultural University,2008,39(9). |
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| Authors: | CHEN Zhongguang ZHANG Guihong XIA Chunli LI Baoxian LI Yijing |
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| Institution: | CHEN Zhongguang,ZHANG Guihong,XIA Chunli,LI Baoxian,LI Yijing(College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China) |
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| 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. |
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| Keywords: | recombinant canine interferon-γ renaturaton by dilution aggregation suppressors activity specific bioactivity inclusion body |
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