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目的普通犬血浆和高免血清分别是白蛋白和特异性免疫球蛋白分离纯化的原材料,针对不同的血浆确立相应的络合条件和解离条件,以分离纯化白蛋白和丙种球蛋白。方法血浆按1∶1体积加入利凡诺溶液,用2%Na2CO3调pH值,使犬血浆与利凡诺进行充分络合;上清液两次络合后解离,获得丙种球蛋白;沉淀解离获得白蛋白。分别对上清液、两次解离液进行蛋白浓度、纯度的测定。结果确定pH 8.5,室温络合2 h,利凡诺浓度在2.3%时为白蛋白的最佳络合条件;pH 6.0,室温络合2 h,利凡诺浓度2.3%为丙种球蛋白最佳络合条件。0.6%氯化钠,0.25%辛酸钠,pH 1.37为白蛋白解离最佳条件;0.85%氯化钠,1.3%甘氨酸,pH 6.8为丙种球蛋白的最佳解离条件。结论利用利凡诺方法,可以成功分离和纯化犬白蛋白和丙种球蛋白。
Objective Normal dogs plasma and serum-free supernatant are albumin and specific immunoglobulin separation and purification of raw materials for different plasma to establish the appropriate complexation conditions and dissociation conditions, in order to separate and purify albumin and gamma globulin. Methods The plasma was dissolved in 1: 1 volume of rivanol solution, and the pH was adjusted with 2% Na2CO3 to fully complex the plasma of the dog with rivanol. The supernatant was dissociated twice after complexation to obtain gamma globulin. The precipitate Dissociate to get albumin. Respectively, the supernatant, two dissociation solution protein concentration, purity determination. The results showed that when pH was 8.5, room temperature complexed for 2 h and the rivanol concentration was 2.3%, the best complexing condition of albumin was pH 6.0, and the concentration of rivanol was 2.3% Complex conditions. 0.6% sodium chloride, 0.25% sodium octanoate, pH 1.37 for the best conditions for albumin dissociation; 0.85% sodium chloride, 1.3% glycine, pH 6.8 for gamma globulin optimal dissociation conditions. Conclusions Canine and gamma globulin can be successfully isolated and purified using the Rivanol method.