摘要
目的应用免疫亲和层析技术快速纯化猪瘟病毒抗体。方法将猪瘟病毒接种于猪传代肾细胞PK-15中培养,收获并纯化猪瘟病毒,将其与环氧氯丙烷活化的Sepharose-4B偶联,制备免疫亲和层析柱,从猪瘟病毒高免疫猪血清中纯化猪瘟病毒抗体,分别应用SDS-PAGE和ELISA进行抗体纯度和活性检测。结果自制免疫亲和层析柱的偶联率为0.36 mg抗原/g载体;纯化的猪瘟病毒抗体纯度高,活性强,抗体回收率占猪血清总蛋白的2.45%。结论已建立了成本低、可快速有效纯化猪瘟病毒抗体的免疫亲和层析法。
Objective To purify the antibody against classical swine fever virus by immunoaffinity chromatography.Methods Classical swine fever virus was inoculated to subcultured pig kidney PK-15 cells and incubated.The harvested virus was purified and coupled with epichlorohydrin-activated Sepharose-4B to prepare immunoaffinity chromatographic column by which the antibody against classical swine fever virus was purified from hyperimmune sera of pigs and determined for purity and activity by SDS-PAGE and ELISA respectively.Results The coupling rate of prepared immunoaffinity chromatography column was 0.36 mg antigen / g carrier.The purified antibody against classical swine fever virus showed high purity and activity,of which the recovery rate of 2.45% of total serum protein.Conclusion A low-cost,rapid and effective immunoaffinity chromatographic method for purification of antibody against classical swine fever virus was successfully developed.
出处
《中国生物制品学杂志》
CAS
CSCD
2011年第10期1233-1235,共3页
Chinese Journal of Biologicals
基金
暨南大学自然科学基金(109057640073)
关键词
免疫亲和层析
猪瘟病毒
高免疫猪血清
Immunoaffinity chromatography
Classical swine fever virus
Hyperimmune pig serum
