CAJ | 학술논문

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∶32000),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.
목적:제비고효개、고특이성적항인광억소 C 계란황 IgY 항체,병대기기본특성진행분석화감정.방법:이인광억소 C 위항면역산단적라만계,채용수희석-염석법제취급순화 IgY 항체,채용단백질정량、SDS-PAGE、West?ern 인적화 ELISA 법대 IgY 항체진행분석화감정.결과:면역후14 d 즉가종계관혈중검측출항광억소 C 적특이성항체,항체효개재28 d 체최고봉(1∶32000),병가유지2개월이상;수집고효개시적면역계단,제비계란황항체 IgY;환성 SDS-PAGE 현시항체 IgY 위상대분자질량분별위65×103화21×103적2조대,항체순도가체92%,득솔위매개계단36.5 mg,항체검출민감도위15.63 ng/mL;Western 인적증명해항체구유고도특이성.결론:제비료항광억소 C 적고효개、고특이성 IgY 항체.
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 cystatin C could be detected from the blood serum of chicken at 14 d post-immunization. The potency of antibody against cystatin 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 92% and the sensitivity detection for cystatin 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.