人乳头瘤病毒重组蛋白疫苗酶联免疫检测法的建立及其应用

目的建立人乳头瘤病毒重组蛋白疫苗(HPV16 L2E6E7)酶联免疫的检测方法,用于疫苗发酵过程中的重组蛋白表达的监控。方法用常规方法制备兔抗HPV16 L2E6E7多克隆抗体作为包被抗体,商品化小鼠抗HPV16 E7单克隆抗体为检测抗体,夹心ELISA方法检测HPV16 L2E6E7抗原参考品,建立抗原标准曲线,确定线性范围及检测限,同时验证该方法的特异性。结果建立了检测HPV16 L2E6E7抗原含量的夹心ELISA方法,抗原参考品系列浓度在19.53~1 250 ng·m L-1内有很好的线性(R2>0.99)和回收率(90%~110%);特异性好,不受发酵液中宿主菌蛋白的干扰。结论建立了人乳头瘤病毒重组蛋白疫苗重组抗原含量的测定方法,为该疫苗的发酵工艺的质控提供了有效技术手段。

Establishment and Application of ELISA Method for HPV16 L2E6E7 Vaccine

OBJECTIVE To establish a quantitative assay method for determining the titer of HPV16 L2E6E7 vaccine during its development and manufacturing. METHODS By using a conventional method, rabbit anti-HPV16 L2E6E7 polyclonal antibodies were generated and used together with a commercially available mouse anti-HPV16 E7 monoclonal antibody to establish a sandwich ELISA method for HPV16 L2E6E7 vaccine. The linearity, sensitivity, and specificity of the method were evaluated. RESULTS A specific and sensitive ELISA method for assaying HPV16 L2E6E7 vaccine quantitatively was developed. Good linearity was shown within the detection range of 19.53-1 250 ng·mL^-1(R²>0.99), and the recovery rate was 90%-110%. The method showed strong sensitivity and high specificity with no interference by host cell proteins of E. coli. CONCLUSION A quantitative ELISA method is established for HPV16 L2E6E7 vaccine. It offers a useful technical tool for quality control of the vaccine during its fermentation process.