抗氯霉素卵黄抗体的制备及其分离纯化

Preparation and Purification of Immunoglobulin Yolk against Choramphenicol

将合成的氯霉素(CAP)与牛血清白蛋白(BSA)的偶联物(CAP-BSA)作为免疫抗原注射到母鸡体内得到氯霉素卵黄抗体(IgY).比较了不同的提取方法,用优化的辛酸-硫酸铵沉淀法从卵黄液中初步提取卵黄抗体.选用3,3′-二氨丙基亚胺(DADPA)作为亲和凝胶与配基氯霉素之间的“间隔臂”,合成了分离纯化抗氯霉素特异性IgY的亲和层析柱.将IgY粗品经过亲和层析柱,得到的特异性IgY的抗体活性提高了10倍,收率约为3.3%.

Yolk immunoglobulin （IgY） was obtained by injecting chloramphenicol （CAP） bound to bovine serum albumin （BSA） into the hens. A primary purification was conducted according to several kinds of methods, such as PEG and ammonium sulphate precipitations. A relatively high purity and specific activity of IgY were obtained by an optimized method of capralic acid-ammonium sulphate precipitation, and a plenty ofphospholipid was well separated, which led to benefiting to store for a long time. Using bis （3-aminopropyl） amine as a spacer between the gel and the CAP ligand, a separation material was synthesized for affinity chromatography to purify the specific IgY against CAP. The specific IgY was about 3.3% in the total IgY. The antibody activity was increased by 10 times after purification by affinity chromatography.