重组人心肌肌钙蛋白I纯化方法的对比

目的：对比两种重组人心肌肌钙蛋白 I（rhcTnI）纯化方法，获取稳定的 rhcTnI，促进心肌肌钙蛋白（cTnI）诊断标准化的研究。方法超声破碎工程菌获取 rhcTnI 包涵体，经2％ Tritonx-100，2 mol/L 脲洗涤后溶解在8 mol/L 脲中，分别经 CM-FF 柱上复性和稀释复性纯化 rhcTnI，对比两种方法纯化 rhcTnI 的获得率及其产物在4、-20、-80℃及冻干条件下的稳定性，确立高效获取稳定的 cTnI 的纯化方法。结果0．1 g 湿重包涵体经 CM-FF 柱上复性和稀释复性的获得率分别为26．8％和18．9％。4、-20、-80℃及冻干条件下，CM-FF 下柱上复性获得的 rhcTnI 稳定，并且柱上复性纯化周期短，效率高。结论 CM-FF 柱上复性要比稀释复性纯化 rhcTnI 高效、稳定。

Comparison in purification methods of the recombinant human cardiac troponin I

Objective To compare the two kinds of purification method for purifying recombinant human cardiac troponin I（cT-nI）to obtain the stable cTnI and promote the study of cTnI diagnosis standardization.Methods The cTnI inclusion body was ob-tained by the ultrasonic broken engineering,after washing by 2% Tritonx-100,2M urea,dissolved in 8M urea,then purified by the column refolding on CM-FF and the dilution refolding respectively.The cTnI yields were compared between the two kinds of meth-od and the stability at 4 ℃,20 ℃,-80 ℃ and on the freeze-dried condition was compared.Then the purification method to effi-ciently obtain the stable cTnI was established.Results The protein about 2 mg and 1.4 mg could be obtained by CM-FF on the col-umn refolding and the dilution refolding from 0.1 g of wet inclusion body,respectively.The former method had the short cycle and high efficiency.The cTnI purified by the column refolding on CM-FF was more stable at 4 ℃,20 ℃,-80 ℃ and on the freeze-dried condition.Conclusion The column refolding on CM-FF is more stable and highly efficient in purification of cTnI than the dilution refolding.