建立双抗体夹心法ELISA定量检测戊型肝炎p179疫苗

发布时间：2018-07-15 18:09

【摘要】：目的:建立双抗体夹心法ELISA检测HEV抗原,用于定量检测戊型肝炎p179疫苗生产过程中初制品、半成品和疫苗成品。方法:将单克隆抗体3G1、5G5、1G10、3A10和4E9分别进行辣根过氧化物酶标记,通过竞争ELISA检测确定最佳抗体配对用于建立双抗体夹心法ELISA,对p179生产过程中所得制品进行抗原定量检测。结果:5种单克隆抗体可以分成两组,分别针对p179疫苗抗原上的两种不同抗原表位,选择3G1为包被抗体、4E9为酶标抗体建立的双抗体夹心法ELISA具有良好的敏感性和特异性,对p179抗原检测的最低浓度可至15.6 ng·ml-1。该方法能够有效检测p179疫苗生产过程中的初制品、半成品和疫苗成品的抗原含量,反映抗原纯化各步骤的得率。结论:建立的双抗体夹心法ELISA可作为p179疫苗生产过程中的抗原检测方法,这有助于定量检测p179疫苗生产过程中的抗原成分。
[Abstract]:Objective: to establish a double antibody sandwich Elisa for the quantitative detection of HEV antigen in the production of hepatitis E p179 vaccine. Methods: horseradish peroxidase was labeled with monoclonal antibody 3G1G5G10A10 and 4E9 respectively. The best pair of antibodies was determined by competitive Elisa to establish the double antibody sandwich method ELISA.The products obtained in the production of p179 were detected quantitatively by antigen. Results five kinds of monoclonal antibodies could be divided into two groups. The double antibody sandwich Elisa with 3G1 as the coated antibody and 4E9 as the enzyme labeled antibody had good sensitivity and specificity for the two different epitopes of p179 vaccine antigen. The lowest concentration of p179 antigen was 15.6 ng / ml ~ (-1). This method can effectively detect the antigen content of the primary, semi-finished and vaccine products in the production of p179 vaccine, and reflect the yield of each step of antigen purification. Conclusion: the double antibody sandwich Elisa can be used as an antigen detection method in the production of p179 vaccine, which is helpful to quantitatively detect the antigen components in the production of p179 vaccine.
【作者单位】：东南大学医学院;长春生物制品研究所有限责任公司;
【基金】：国家863疫苗重点项目(2006AA02A235) 东南大学国家大学生创新训练计划项目(1210286097)
【分类号】：R512.6

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