人补体片段C5a蛋白的可溶性表达、纯化及生物学活性检测

摘要

Objective:To prepare the soluble human complement fragment C5a protein and measure its bioactivity.Methods:The C5a fusion protein was induced to express by IPTG and then digested by enterokinase,purified by Ni2+-NTA column.By the neutrophil chemotaxis,respiratory burst and producing lactoferrin experiment,the bioactivity of C5a protein was be detected.Results:The C5a fusion protein is soluble expression,high purity C5a protein is obtained after the fusion protein be digested by enterokinase.The C5a protein can lead to human neutrophil chemotaxis,cause human neutrophil respiratory burst and induce human neutrophils produce lactoferrin.Conclusion:The human complement fragment C5a protein with bioactivity can be expressed in soluble form.The study laid a foundation for the further clinical research and application.%目的：：为得到可溶性表达的人补体片段C5a蛋白,并检测其生物学活性。方法：用IPTG对C5a融合蛋白进行诱导表达,表达产物经肠激酶酶切后用Ni2+分离柱纯化得到C5a蛋白,通过对中性粒细胞的趋化作用、呼吸爆发、产生乳铁蛋白等实验检测C5a蛋白的生物学活性。结果：成功获得可溶性表达的C5a融合蛋白,通过肠激酶酶切后得到高纯度的C5a蛋白,C5a蛋白对人中性粒细胞具有明显的趋化作用,能引起人中性粒细胞的呼吸爆发并能诱导人中性粒细胞产生乳铁蛋白。结论：可溶性表达了具有生物学活性的人补体片段C5a蛋白,为进一步的临床研究和应用奠定了基础。