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鸡血清载脂蛋白AⅠ的分离纯化及其抗血清的制备

Separation and purification of apolipoprotein AⅠ from chicken serum and preparation of its specific antiserum

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【作者】 李浩棠张彩英曹华斌郭小权俞海峰胡国良

【Author】 LI Hao-tang, ZHANG Cai-ying, CAO Hua-bin, GUO Xiao-quan, YU Hai-feng, HU Guo-liang(College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045,China)

【机构】 江西农业大学动物科学技术学院江西农业大学动物科学技术学院 江西南昌330045江西南昌330045江西南昌330045

【摘要】 用磷钨酸沉淀制备鸡血清HDL,乙醇丙酮混合液(体积比为1∶1)脱脂后采用DEAESepharoseCL6B离子交换柱层析法分离鸡血清apoAⅠ,分离获得的apoAⅠ在尿素SDSPAGE电泳中呈现单一条带,其分子质量约为27788u。将提取的apoAⅠ对新西兰雄兔进行多次免疫,制备得兔抗鸡apoAⅠ抗血清,其效价为1∶16。在免疫电泳和双向免疫扩散试验中抗血清均只呈现1条清晰沉淀带。试验结果表明,用磷钨酸沉淀法制备兔抗鸡apoAⅠ抗血清,过程简便,时间短且分离效果好。

【Abstract】 The HDL of laying hen was isolated and purified from pooled serum by precipitating with phosphotungstic acid. The apolipoprotein AⅠ was obtained from HDL by delipitating with acetone-ethanol(Ψ=1)and purified by ion exchange chromatography with DEAE sepharose CL-6B. The identification by 12.5% urea-SDS-PAGE showed that the purified apolipoprotein AⅠ have only one band and its mole- cular weight is 27788u. The specific antiserum was prepared by immunizing New Zealand male rabbit with the obtained protein for several times. The titer of the prepared specific antiserum was 1∶16, and the specific antiserum showed a single sedimentary band on immune electrophoresis and double immunodifusion. The results showed that the preparation of the rabbit-anti chicken antiserum by the phosphotungstic acid precipitation method was simple, saved times and could achieve good results.

【基金】 国家自然科学基金资助项目(30160064)
  • 【文献出处】 中国兽医科技 ,Chinese Journal of Veterinary Science and Technology , 编辑部邮箱 ,2005年07期
  • 【分类号】S852.23
  • 【被引频次】3
  • 【下载频次】240
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