中文摘要：

为建立猪睾丸细胞疫苗生物制品中宿主细胞蛋白残留量的ELISA检测方法,制备了猪睾丸细胞多克隆抗体,优化了间接ELISA试验条件,建立了ELISA检测方法。Western-blot结果显示,制备的猪睾丸细胞蛋白多克隆抗体具有良好的特异性,抗体经过纯化后效价达1∶10 000。猪睾丸细胞蛋白的质量浓度为25-400ng/mL时线性关系良好。结果表明,用猪睾丸细胞蛋白建立的间接ELISA定量检测方法具有良好的线性关系、特异性和重复性。

英文摘要：

To establish an ELISA for detection of host cell proteins（HCP）of porcine testis in biologicals,the soluble proteins were prepared from swine testical（ST）cells,and then were injected into rabbits to produce high titers of polyclonal antisera.The titers and specificity of rabbit sera were determined by ELISA and Western-blot,respectively.Using the purified polyclonal antibodies from rabbits,an ELISA was established for determining host cell proteins of swine testis in biologicals.In result,the titers were more than 1∶10 000 and the polyclonal antibodies against ST HCP prepared had a good specificity and reactivity.The polyclonal antibodies have no cross-reactions with DMEM,MEM,newborn bovine serum,fetal bovine serum,and horse serum,respectively.The ELISA method developed had good specificity,sensitivity,and reproducibility and the detection limit was 25 ng per milliliter.The linear range of the standard curve is between 25 to 400ng per milliliter and the correlation coefficient was 0.999.In conclusion,aquantitative ELISA was established for detecting swine testical cell proteins,which provides a reliable method for detection of the host cell proteins in biological products.