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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ELISA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60 min,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 To prepare the yolk antibody against Cnidium serrata and its stability. Methods The primary and secondary immunization of healthy Lai-han hen was carried out by using the white-leaf bamboo green snake venom. The yolk antibody was purified by the combination of water extraction, salting-out and thionylchromatography. The indirect ELISA Its potency and stability were evaluated. Results After immunization, the yolk in chicken yolk could produce a higher level of antibodies to C. albiflorus and maintain it for 10 weeks. The IgY purified by three-step method showed a single band on SDS-PAGE. It maintained stable activity at the temperature range of 20 ℃ ~ 65 ℃ and the pH value ranged from 4 ~ 12. And the activity decreased slightly after trypsin treatment for 60 min. Conclusion Antibodies against white-leaf bryozoan snake venom obtained by this method have the advantages of simple preparation and good stability.