| 虎血清免疫球蛋白（IgG）的纯化及纯度、活性鉴定 |
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| 引用本文： | 刘先菊,;杨帆,;林树柱,;全雄志,;夏咸柱,;张成林,;秦川,;张连峰.虎血清免疫球蛋白（IgG）的纯化及纯度、活性鉴定[J].中国实验动物学杂志,2007(11):637-640. |
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| 作者姓名： | 刘先菊 ;杨帆 ;林树柱 ;全雄志 ;夏咸柱 ;张成林 ;秦川 ;张连峰 |
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| 作者单位： | [1]中国医学科学院实验动物研究所北京协和医学院比较医学中心,北京100021; [2]军事医学科学院军事兽医研究所,长春130062; [3]北京市动物园兽医院,北京100044 |
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| 基金项目： | 社会公益研究专项（2005DIA1J095）. |
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| 摘 要： | 目的 建立高纯度、高活性的虎血清IgG纯化方法。方法 用饱和硫酸铵沉淀虎血清得到IgG粗品;结合Hitrap Protein A亲和层析预装柱及阴离子交换层析法对粗品IgG进一步分离纯化,采用PAGE电泳和Western-Blot免疫印迹法鉴定IgG纯度和免疫活性。结果 80 mL虎血清亲和纯化得到84 mg IgG,阴离子交换层析纯化得到30 mg虎的IgG纯品。结论 建立了简便快速、纯度高、活性好的虎血清IgG的分离纯化方法,为虎血清IgG二级抗体的制备提供了高纯度、活性好的一级抗体免疫原。
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| 关 键 词： | 亲和层析 阴离子交换层析 老虎 IGG |
Purification and Identification of IgG from Tiger Serum |
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| Institution: | LIU Xian-ju, YANG Fan, LIN Shu-zhu, QUAN Xiong-zhi, XIA Xian-zhu, ZHANG Cheng-lin,QIN Chuan, ZHANG Lian-feng ( 1. Institute of Laboratory Animal Science, Chinese Academy of Medical Science, Comparative Medicine Center, Peking Union Medical College, Beijing 100021, China; 2. Institute of Military Veterinary, Academy of Military Medical Sciences, Changchun 130062,China; 3. Amimal Hospital Beijing Zoo, Beijing 100044, China) |
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| Abstract: | Objective To establish a method for purification of IgG from tiger serum with high purity. Methods The tiger serum was subject to precipitation with ammonium sulfate and then purified by Protein-A Affinity Chromatography and anion exchange chromatography, The purity of IgG was identified by SDS-PAGE and Western-Blot methods. Results Eighty-four mg IgG was purified from 80 mL tiger serum using Protein-A Affinity Chromatography and Thirty mg IgG was further purified from the 84mg affinity purification IgG using anion exchange chromatography. Conclusion The method was established for purification of tiger lgG, and provide lgG antigens for the production of polyclonal rabbit anti-tiger-lgG secondary antibody. |
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| Keywords: | Affinity chromatography Anion exchange chromatography Tiger IgG |
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