抗幽门螺杆菌蛋黄免疫球蛋白IgY的制备及生物活性的研究

A Study on Preparation of Anti-helicobacter Pylori Egg Yolk IgY and Its Biological Activity

目的:从免疫了幽门螺杆菌(Hp)抗原的母鸡蛋黄中提取抗Hp-IgY。方法:用超声破碎的Hp抗原免疫21周龄罗曼产蛋母鸡,取免疫后所产鸡蛋卵黄,利用水稀释法加硫酸铵盐析方法,提纯IgY,ELISA间接法检测抗体效价,Bradford法检测抗体蛋白含量,SDS-PAGE法检测其相对分子质量及纯度。结果:母鸡初次免疫后第7天,抗体效价即渐渐升高,约第45天达到高峰;蛋黄抗体经纯化后,抗Hp-IgY蛋白纯度可达90%以上,含量为6.25 g/L。结论:用超声破碎的Hp抗原免疫母鸡,盐析法提纯免疫母鸡产蛋黄中的抗Hp-IgY,可获得高纯度及高效价的抗Hp的特异性蛋黄抗体。

Objective： To extract and purify anti-helicobacter pylori IgY from egg yolk of hens immunized by Helicobacter pylori（HP）.Methods： HP antigen subjected to ultrasonication was used to immune 21 weeks old Roman egg production hens.IgY in immunized yolk was isolated by water dilution method combined with ammonium sulfate salting out method.Its titer was detected by indirect ELISA.The protein amount was measured with Braford method.The purity and molecular weight of IgY were measured with SDS-PAGE method.Results： Seven days after the first immunization,the IgY titer gradually increased,and reached the peak on the 45th days.After purification,the purity of IgY was more than 90%,and the protein content was 6.25 g/L.Conclusions： In this way,anti-HP IgY with high purity and titer has been prepared.