在制备尖吻蝮蛇毒出血毒素抗血清中不同凝胶染色方法的比较  

Comparative studies by different dyeing methods on antiserum preparation against hemorrhagins from Agkistrodon acutus venom

作　　者：韦传宝[1] 曹佳敏[1] WEI Chuan-Bao CAO Jia-Min(Biological and Pharmaceutical Engineering College, West Anhui University, Liuan 237012, China)

机构地区：[1]皖西学院生物与制药工程学院,六安237012

出　　处：《中国免疫学杂志》2016年第12期1793-1796,共4页Chinese Journal of Immunology

基　　金：安徽自然科学基金项目(1408085MC43);安徽省教育厅自然科学重点项目(KJ2015A454;KJ2013A264)

摘　　要：目的:寻找在制备出血毒素抗血清中电泳后染色凝胶的好方法。方法:按照前人的方法从皖南产尖吻蝮蛇毒中初步分离3种出血毒素:Aa HⅠ、Aa HⅡ和Aa HⅣ并获得3种粗提取的出血毒素,用制备电泳的方法进一步纯化Aa HⅠ、Aa HⅡ和Aa HⅣ,对每种出血毒素,配制6块制备电泳板并进行聚丙烯酰胺凝胶电泳,用6种不同的染色方法分别染色,将含出血毒素的凝胶切下,磨细后直接免疫注射小鼠并获得抗血清,用ELISA检测和中和出血毒素出血活性两种方法检测抗血清的质量。结果:不同染色方法制备的抗血清中有效Ig G浓度不同,用无毒型蛋白质快速染色试剂盒染色制备的抗血清有效Ig G浓度最高。结论:用无毒型蛋白质快速染色试剂盒染色是制备抗血清最好的染色方法。Objective：In order to look for a good method for preparation of hemorrhagin antiserum. Methods： Three kinds of hemorrhagins including AaH Ⅰ , AaH Ⅱ , and AaH Ⅳ were purified from Agkistrodon acutus venom according to predecessors＇s methods and crude AaH Ⅰ , AaH Ⅱ and AaH IV were obtained. Preparation electrophoresis was used to purify AaH Ⅰ ,AaH Ⅱ and AaH Ⅳ further. As for an hemorrhagin, six different dyeing methods were used to dye PAGE gel and the gel contained hemorrhagin was obtained respectively. The ground gel contained hemorrhagin was used to immune mice and its antiserum was obtained. Antiserums quality was tested through ELISA test and neutralization of the hemorrhagic activities of corresponding hemorrhagin. Results： Effective IgG concentration in different antiserum was different and effective IgG made through non toxic type protein fast stain reagent kit was higher than others. Conclusion： Non toxic type protein fast stain reagent kit is the best dyeing method among the six dyeing methods.

关 键 词：尖吻蝮蛇 出血毒素 抗血清制备 染色法 质量检测 

分 类 号：R991[医药卫生—毒理学]