利用水稀释反复冻溶法、氯仿萃取法与冷乙醇沉淀法相结合进行鸡新城疫卵黄抗体提取的试验研究

Study on Extraction of Chicken ND-IgY Combining with Water Dilution Repeated Freezing and Dissolving Method,Chloroform Extraction and Cold Ethanol Precipitation

本研究采用免疫学技术和物理化学方法,通过优化提取条件,改良提取工艺,建立了一种用水稀释反复冻溶法、氯仿萃取法与冷乙醇沉淀法相互结合提取鸡卵黄抗体的方法。结果显示:用5倍体积、pH5.0的稀释液稀释蛋黄,在-20℃经三次反复冻融、上清液再用同等剂量的氯仿萃取、40%的冷乙醇沉淀,提取的鸡新城疫卵黄抗体具有液体清亮,杂质含量少,抗体效价高的特点。

An extraction method of egg yolk IgY combining with water dilution repeated freezing and dissolving Method, chloroform extraction and cold ethanol precipitation was established using immunological techniques and physical chemical method through the optimization of extraction conditions and improved extraction proeess. The results showed that after diluted egg yolk with 5 times volume diluents （pH5.0） and passed by three times of repeated freezing and thawing, extracted by equal doses chloroform and then precipitated with 40% cold ethanol at -20℃, the extraction liquid of the chicken ND--IgY has a clear liquid characteristics with low impurity content and high antibody titer.