抗胱抑素C鸡卵黄IgY抗体的制备及鉴定  

Preparation and Characterization of IgY Antibody Against Cystatin C

作　　者：傅龙章[1] 李敏惠[2] 潘克俭[1] 邹珂珂[1] 杨菌[1] 李艳超[1] 黄杰[1] 叶启迪[1] 马玲玲[1] 杨平[3] 

机构地区：[1]成都医学院生物医学系,四川成都610083 [2]成都医学院科研实验中心,四川成都610083 [3]成都医学院生物化学教研室,四川成都610083

出　　处：《生物技术通讯》2013年第2期222-224,共3页Letters in Biotechnology

基　　金：四川省教育厅青年基金(10ZB095);成都医学院创新实验项目(CX201108);成都医学院横向课题(10H018);成都医学院科研基金(CYZ11-005)

摘　　要：目的:制备高效价、高特异性的抗人胱抑素C鸡卵黄IgY抗体,并对其基本特性进行分析和鉴定。方法:以人胱抑素C为抗原免疫产蛋的罗曼鸡,采用水稀释-盐析法提取及纯化IgY抗体,采用蛋白质定量、SDS-PAGE、West ern印迹和ELISA法对IgY抗体进行分析和鉴定。结果:免疫后14 d即可从鸡冠血中检测出抗胱抑素C的特异性抗体,抗体效价在28 d达最高峰(1∶32 000),并可维持2个月以上;收集高效价时的免疫鸡蛋,制备鸡卵黄抗体IgY;还原性SDS-PAGE显示抗体IgY为相对分子质量分别为65×103和21×103的2条带,抗体纯度可达92%,得率为每个鸡蛋36.5 mg,抗体检出敏感度为15.63 ng/mL;Western印迹证明该抗体具有高度特异性。结论:制备了抗胱抑素C的高效价、高特异性IgY抗体。Objective： To prepare high potent and specific IgY antibody against cystatin C. Methods： Hens were immunized with cystatin C as antigen and the antibody was purified through the water dilution and sodium sul- phate salting-out methods from eggs. The antibody was characterized through methods including SDS-PAGE, West- ern blot and ELISA. Results： Specific IgY antibody against eystatin C could be detected from the blood serum of chicken at 14 d post-immunization. The potency of antibody against eystatin C could reach the peak 1：32 000 at 28 d post-immunization and maintain high production in subsequent 2 months. The IgY antibody was purified when valence of antibody was at the peak. Reduced SDS-PAGE molecular weights of IgY were of around 65 and 21 kD respectively. Normally, 36.5 mg IgY could be isolated from each egg yolk, the purity of IgY was 920/o and the sensitivity detection for eystatin C was 15.63 ng/mL. Western blot showed the antibody has good specificity. Conclusion： High potent and specific IgY antibody against cystatin C which may be a novel and useful diagnosis reagent was successfully prepared in this study.

关 键 词：IGY抗体 胱抑素C 鸡卵黄 

分 类 号：R392.1[医药卫生—免疫学]