抗白唇竹叶青蛇毒卵黄抗体(Ig Y的制备及其稳定性研究  

Study on the development of antibody(IgY) against Trimeresurus albolabris venom from Hen Egg Yolk and its Physicochemical Stability

作　　者：段海龙[1] 余晓东[1] 邓骛远[2] 杨宪[1] 刘刚[1] 王文文 和七一[1] 

机构地区：[1]重庆市动物生物学重点实验室、重庆市生物活性物质工程研究中心、教育部活性物质生物技术工程研究中心、重庆师范大学生命科学学院,重庆400047 [2]宜宾学院生物研究所,四川宜宾644000

出　　处：《现代预防医学》2014年第6期1074-1077,共4页Modern Preventive Medicine

基　　金：重庆市自然科学基金(cstc2011jjA10100);大学生创新创业训练计划项目(201210637003)

摘　　要：目的制备抗白唇竹叶青蛇毒卵黄抗体，并对其稳定性进行研究。方法采用白唇竹叶青蛇毒对健康莱杭母鸡进行初次免疫和两次加强免疫；通过水提法、盐析法和嗜硫色谱法相结合对抗白唇竹叶青蛇毒卵黄抗体进行纯化；采用间接EL｜SA法对其效价及稳定性进行评价。结果免疫后鸡卵黄中可以产生较高水平的抗白唇竹叶青蛇毒抗体，并可维持至10周；经三步法纯化得到的抗体IgY在SDS—PAGE上呈单一条带。其在20℃～65℃及pH值为4～12范围内都保持稳定的活性；而且经胰蛋白酶处理60rain，其活性降低趋势不明显。结论此法获取的抗白唇竹叶青蛇毒抗体具有制备简单、稳定性好的优点。Objective This work was to develop antibody （IgY） against the venom of Trimeresurus a!bolabris from hen egg yolk and to study its stabilities. Methods The venom of Trimeresurus cdbolabris was injected to leghorn hens at the initial immunization and two booster immunizations. The IgY was purified sequentially with water extraction, salt precipitation and thiophilic chromatography. ELISA was used to indirectly evaluate the titer and stability of IgY. Results IgY obtained after immunizing the hens showed high titer which could last for 10 weeks or so. The IgY presented only one band on SDS-PAGE, kept active in the rang of 20 - 65℃, or in the rang of pH 4 -12 at 37℃ for two hours. No significant reduction in activity was observed after it was mixed with trypsin for 1 hour. Conclusion This method for developing IgY against the venom of T. albolabris from hen egg was simple, convenient and reli- able since the obtained IgY antibody exhibited good stability under adversed conditions.

关 键 词：白唇竹叶青蛇 卵黄抗体 稳定性 酶联免疫吸附测定 

分 类 号：Q-33[生物学]