牛血清蛋白酶联免疫检测试剂盒的研制及验证

为研制酶联免疫试剂盒以检测病毒性疫苗中残余牛血清蛋白(BSP)含量,制备高效价高纯度的兔抗BSP多克隆抗体作为包被抗体和酶标抗体,建立了ELISA双抗体夹心法并组建试剂盒,通过标准剂量曲线可对样品中所含BSP、BSA及B-IgG进行定量,经验证该方法标准曲线线性范围内r≥0.98,对BSP的检测限量为3ng/ml;分别检测5、10、20ng/ml含量的BSP时,试验内(n=12)和试验间(n=3)测定的变异系数在3.71%到7.29%之间,回收率在93.4%~106.3%,未见该方法与人血清白蛋白、卵清蛋白以及疫苗复合保护剂之间有交叉反应。该法敏感度高,准确性、重复性和稳定性好,可用于疫苗牛血清残余蛋白的质量控制。

Development and validation of ELISA kit on detecting residual bovine serum proteins in viral vaccines

To measure the contents of residual bovine serum proteins(BSP)on developing ELISA kit.ELISA method was established and ELISA kit was developed by coating high titer antibody in the plate purified from rabbit serum and labelling the same antibody with HRP as detection antibody,which can detect bovine serum proteins,bovine serum albumin and bovine IgG by the standard dose curve.The detection quanta was as low as 3ng/ml and the precision was preferable because the coefficients of variation of the results detecting the BSP standards of 5,10,20ng/ml concentrations were from 3.71% to7.29%,and the recovery from 93.4% to 106.3% indicated measuring results had return accuracy.There was no cross reaction observed when detecting human serum albumin,ovalbumin and vaccine protectins.The method could be used for data quality monitoring of residual bovine serum albumin in vaccines of our company.