犬血清IgG的纯化、抗体制备及其鉴定

[目的]探讨犬血清IgG的纯化方法。[方法]采用硫酸铵盐析法结合SephadexG-200凝胶柱层析提取纯化犬血清IgG,利用SDS-PAGE电泳和免疫印迹法鉴定所得产品的纯度和免疫活性。[结果]获得的纯化犬血清IgG的SDS-PAGE图谱中存在2条蛋白条带,表明获得了高纯度的犬血清IgG;重链分子量为58kD,轻链分子量为27kD,IgG分子量为170kD。特异性检测发现犬血清IgG重链和轻链都能与鼠抗犬血清IgG抗体结合,表明纯化犬血清IgG具有免疫原性和反应原性。[结论]硫酸铵盐析法结合SephadexG-200凝胶柱层析可获得纯度高、活性好的纯化犬血清IgG,为犬血清IgG相关的免疫学试验提供了高质量的免疫材料。

Purification and Antibody Preparation of Canine Serum IgG and its Identification

[Objective] The research aimed to explore the purification method of canine serum IgG.[Method] The purity and immune activity of canine serum IgG were identified by SDS-PAGE electrophoresis and western blot after extracted and purified by ammonium sulfate salting-out method combined with SephadexG-200 gel filtration chromatography.[Result] The purified canine serum IgG showed two protein bands in the SDS-PAGE map,which indicated that the obtained canine serum IgG was highly purified.The molecular weight of heavy chains,light chains and IgG were 58 kD,27 kD and 170 kD,resp.Specific detection showed that the heavy chains and light chains of canine serum IgG all could bind with mouse anti canine serum IgG,which indicated that the purified canine serum IgG had immunogenicity and reactiongenicity.[Conclusion] The purified canine serum IgG with high purify and activity were obtained by ammonium sulfate salting-out method combined with SephadexG-200 gel filtration chromatography,which provided high quality immune material for canine serum IgG-related immune experiment.