抗竹叶青蛇毒抗体F(ab′)2的制备  被引量:6

Preparation of F(ab′)2 fragment of Trimeresurus stejnegeri antibody

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作  者:郑颖[1] 张连江[2] 范泉水[1] 邱薇[1] 李作生[1] 

机构地区:[1]成都军区疾病预防控制中心,昆明650032 [2]云南农业大学动物科学技术学院,昆明650201

出  处:《中国药科大学学报》2007年第3期269-272,共4页Journal of China Pharmaceutical University

基  金:全军医学科学技术研究"十五"计划指令性课题资助项目(No.01L009);全军医学科研"十五"计划第二批杰出人才基金课题资助项目(No.04J004)~~

摘  要:目的:制备特异性强、高纯度的抗竹叶青蛇毒抗体F(ab′)2。方法:以竹叶青蛇毒为免疫原免疫马,试血效价达到要求后,从免疫马血浆中先提取IgG,然后酶解、过疏水柱后得到F(ab′)2片段,以免疫扩散实验检测抗体F(ab′)2片段的特异性,以小鼠中和试验测定其中和竹叶青蛇毒的能力。结果:纯化得到的F(ab′)2活性片段纯度可达90%;F(ab′)2冻干品与竹叶青蛇毒在8∶1时出现免疫沉淀线,在20∶1时可保护小鼠100%存活。结论:纯化得到的抗体F(ab′)2活性片段对竹叶青蛇毒具有较好的中和能力。Aim: To prepare the F(ab′)2 fragment of Trimeresurus stejnegeri antibody of high specificity and purity. Methods: IgG was extracted from the horse plasma immunized by the venom of Trimeresurus stejnegeri, then IgG was subjected to pepsin-digest, ammonium sulfate precipitation and chromatograph on hydrophobic column to obtain the F(ab′)2 active fragment. The specificity of antivenin was traced by way of immunodiffusion, and the protective effect was performed on mice in vitro. Results:The results of electrophoresis strip revealed that the purity of F(ab′)2 active fragment was up to 90%. The immune precipitation lines clearly appeared when the proportion of lyophilized antivenin and venom was 8:1; while it was 20:1, the mice are 100% survived in the neutralization test. Conclusion:The obtained antivenin can neutralize the venom efficiently.

关 键 词:竹叶青蛇毒 抗体 F(ab′)2 制备 冻干 

分 类 号:R392[医药卫生—免疫学]

 

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