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目的建立高特异性检测牛血清白蛋白(BSA)竞争ELISA并应用于检测生物制品中残留的BSA。方法采用商品化的BSA作为包被抗原及标准品,筛选高特异性、高纯度的抗BSA单克隆抗体(mAb)作为酶标抗体,建立敏感、特异的竞争ELISA。结果优化了竞争ELISA的实验条件,对BSA的最低检测限为1.0 ng/mL,批内实验变异系数为4.3%~7.0%,回收率为92%~108%。该方法与人血清、马血清、兔血清、鼠血清、鸡血清均无交叉反应。结论该方法特异性强、灵敏度高、重复性好,适用于生物制品中BSA残留以及其他标本BSA的定量检测。
OBJECTIVE: To establish a competitive ELISA for bovine serum albumin (BSA) assay and to detect BSA residues in biological products. Methods Commercially available BSA was used as coating antigen and standard to screen highly specific and high purity anti-BSA monoclonal antibody (mAb) as enzyme-labeled antibody to establish a sensitive and specific competitive ELISA. Results The competitive ELISA conditions were optimized. The detection limit of BSA was 1.0 ng / mL. The coefficient of variation (CV) was 4.3% -7.0% and the recovery was 92% -108%. The method has no cross reaction with human serum, horse serum, rabbit serum, rat serum and chicken serum. Conclusion The method is specific, sensitive and reproducible. It is suitable for the quantitative determination of BSA in biological products and BSA in other specimens.