人C-反应蛋白的分离纯化及初步应用

Isolate Human C-reactive Protein and Preliminary Application

目的:从临床血清样本分离纯化获得人C-反应蛋白(CRP)。方法:运用盐析法、DEAE离子交换层析和亲和层析等方法从临床血清样本中分离纯化出CRP,用于免疫山羊制备多克隆抗体,初步建立免疫透射比浊法测定CRP并进行评价。结果:SDS-PAGE结果显示纯化产物出现相对分子质量约为23000的主带,Western Blotting结果显示纯化产物能与CRP单克隆抗体发生特异性结合,证实纯化产物为CRP,与商品CRP有很好的可比性性。以自提CRP制备出效价高、特异性强的多克隆抗体,运用该抗体建立的CRP免疫透射比浊法具有比较好的性能。结论:成功分离纯化出人CRP,可用于进行生产研究。

Objective：To isolate and purify human C-reactive protein（h-CRP） from clinical serum sample.Methods：Isolated and purified h-CRP from clinical serum sample by salting-out method,ion exchange and affinity chromatography,a goat was immunized with the purified h-CRP for prepare PcAb,preliminarilly established immunoturbidimetry for determining h-CRP,and evaluated it.Results：SDS-PAGE showed that purified product appeared main bound with an apparent molecular weight of 23000,Western Blotting showed that it reacted specifically to CRP McAb,it was ascertained as CRP,the result of it were identical with commercial CRP.Used the purified CRP as immunogen,obtained goat anti-human PcAb with high titer and specificity,Used the PcAb to established immunoturbidimetry for determining h-CRP which had a good performance.Conclusion：The h-CRP was successfully isolated and purified,which can be used for production and research.