摘要
运用 (NH4) 2 SO4沉淀法、冷乙醇沉淀法、硫酸葡聚糖沉淀法对猪血清中的免疫球蛋白Ig进行粗提取 ,然后用透析技术脱盐。结果表明 ,在这三种粗提取方法中 ,以硫酸铵沉淀法为最高 ;硫酸铵沉淀法的最优提取条件是 :pH7.0～ 7.4 ,温度 4 0℃ ,5 0 %的 (NH4) 2 SO4溶液 ,然后改用 33%的SAS可以有效地提取IgG ;透析脱盐时 ,透析膜截留分子量 8,0 0 0～ 12 ,0 0 0 ,透析 72小时 ,用PBS液更换 4～ 6次 ,所得到的透析液基本上不含盐类 ;提取IgG在 2 7℃～ 4 7℃时 ,免疫活力基本没有变化 ,温度从 5 7℃开始变化 ,至 77℃活性基本丧失。当pH低于 4 .5时 。
In this article, the crude extraction of the Immunoglobulin in pig serum is studied by three methods, which include ammonium sulphate precipitate, cold alcohol precipitate and polyglucosan sulphate precipitate, and then the crude extract are desalted by dialysis. The result shows, among these crude extraction methods, ammonium sulphate precipitate is the best, the optimal condition of ammonium sulphate precipitate is that: the pH is from 7.0 to 7.4, the temperature range is from 40℃ and the ammonium sulphate saturation is 50%, it is found that the capture molecular weight of the dialysis membranes is from 8,000 to 12,000, and no salt in the dialysis solution, under the condition of the time lasting for 72 hours, and the PBS refreshing 4～6 times, the antibody activities of resulting product is not affected in the conditions of pH from 7.0 to 7.4, and temperature lower 57℃, but the antibody activities of resulting product is not affected when the pH is lower 4.5, and temperature lower 57℃, and when the temperature is up to 77℃, the antibody activity is almost disappeared completely.
出处
《河南科学》
2002年第5期528-532,共5页
Henan Science
