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›› 2006, Vol. 6 ›› Issue (2): 281-284.

• 生化工程专栏 • 上一篇    下一篇

抗氯霉素卵黄抗体的制备及其分离纯化

吴媛媛,包永明,李晓晖,修志龙   

  1. 大连理工大学环境与生命学院生物科学与工程系
  • 出版日期:2006-04-20 发布日期:2006-04-20

Preparation and Purification of Immunoglobulin Yolk against Choramphenicol

WU Yuan-yuan,BAO Yong-ming,LI Xiao-hui,XIU Zhi-long   

  1. Department of Bioscience & Biotechnology, School of Environmental & Biological Science and Technology, Dalian University of Technology
  • Online:2006-04-20 Published:2006-04-20

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摘要： 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%.

关键词: 氯霉素, 卵黄抗体, 亲和层析

Abstract: 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 of phospholipid 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.

Key words: chloramphenicol, yolk immunoglobulin, affinity chromatography