尖吻蝮蛇出血毒素分离纯化及其抗体在蛇毒鉴别中的应用  

Purified the Hemorrhagic Toxin of Agkistrodon acutus Venom and Its Antibody Applied in Identification Different Venoms

作　　者：郑颖[1] 张绎[1] 叶锋平[1] 聂雪峰[1] 张志晓[1] 邱薇[1] 范泉水[1] 

机构地区：[1]成都军区疾病预防控制中心,昆明650032

出　　处：《中国生物工程杂志》2013年第3期123-129,共7页China Biotechnology

基　　金：国家重大新药创制科技重大专项(2008ZXJ09004-039);全军"十二五"医学科研面上项目(CWS11J280)

摘　　要：目的:纯化尖吻蝮蛇毒出血毒素,制备其特异性抗体,并建立该抗体对多种蛇毒的ELISA检测方法。方法:利用多种层析技术从尖吻蝮蛇蛇毒中分离得到出血毒素,利用该蛋白作为抗原免疫家兔,通过分步盐析和Protein G亲和层析法从免疫血浆中纯化IgG抗体,HRP标记纯化的抗体。用该特异性抗体ELISA法检测蝰科的五种蛇毒。结果:纯化的出血毒素HPLC检测纯度为99.49%,其特异性HRP-IgG抗体能有效从各蝰科蛇毒中鉴别出尖吻蝮蛇毒,与各蝰科蛇毒交叉反应<15%,检测的灵敏度为10ng/ml。结论:利用纯化的抗尖吻蝮蛇蛇毒出血毒素抗体建立的ELISA方法可快速准确地从多种蝰科蛇毒中鉴别出尖吻蝮蛇蛇毒。Aim： To purify the hemorrhagic toxin （HR） of Agkistrodon acutus venom, prepare its special antibody and establish related ELISA method for detection of different snake venoms. Methods： HR was obtained from Agkistrodon acutus venom by several column chromatographs and used as antigen to immunized rabbits. IgG antibody was isolated and purified from immunized rabbit plasma by ammonium sulfate precipitate and affinity chromatography, labeled with HRP. The five venoms from Viperidae were identified by developed ELISA using purified specific igG antibody. Results： The purity of purified HR was up to 99.49 %, ELISA analysis suggested that the specific HRP-IgG antibody can be fast and exactly identified venom of Agkistrodon acutus from other viperidae venoms, and its sensitivity was 10ng/ml, the rate of cross reaction was 〈15%. Conclusion： The developed method of purified HR antibody sandwich ELISA can be quickly and accurately identified venom of Agkistrodon acutus from other kinds of viperidae venoms.

关 键 词：尖吻蝮蛇 出血毒素 分离 抗体 

分 类 号：Q814[生物学—生物工程]