动物过敏原牛血清白蛋白表面等离子共振传感器检测方法的建立

发布时间：2018-04-02 10:13

本文选题：牛血清白蛋白　切入点：动物过敏原　出处：《南京农业大学学报》2017年04期

【摘要】：[目的]建立一种表面等离子共振(surface plasmon resonance,SPR)技术,用于快速、准确、灵敏、定量检测动物过敏原牛血清白蛋白(bovine serum albumin,BSA)。[方法]将鼠抗BSA单克隆抗体偶联于SPR生物传感器芯片表面,监测BSA与抗体结合后芯片表面的变化,并基于SPR技术建立快速检测微量BSA的新方法,确定其检出限和最佳线性范围,并进行重复性验证和初步应用。[结果]抗体的偶联水平为18 000,对BSA具有良好的亲和能力;BSA质量浓度在6.25~400 ng·m L~(-1)范围内,与SPR响应值的变化量呈良好的线性关系(R2=0.992),方法检测限为33.68 ng·m L~(-1);用建立的SPR方法检测不同浓度BSA的日内RSD为1.12%~3.42%,日间RSD为0.84%~3.98%,均小于5%;SPR方法与ELISA商业化试剂盒检测不同市售食品中BSA含量的结果比较,差异不显著(P0.05)。[结论]SPR方法可靠性好、稳定性强,适用于食品中过敏原BSA的高通量实时检测。
[Abstract]:[objective] to establish a surface plasmon resonance technique for rapid, accurate, sensitive and quantitative detection of bovine bovine serum albumin (BSA), an animal allergen. [methods] Mouse anti BSA monoclonal antibody was coupled to the surface of SPR biosensor chip. The changes of chip surface after binding of BSA and antibody were monitored, and a new method for rapid detection of trace BSA was established based on SPR technology, and the detection limit and optimal linear range were determined. The results showed that the coupling level of antibody was 18000, and had good affinity to BSA in the range of 6.25 ~ 400ng / mL ~ (-1). There was a good linear relationship with the response value of SPR. The detection limit of the method was 33.68 ng / mL ~ (-1); the intraday RSD of different concentrations of BSA by the established SPR method was 1.12 ~ (3.42) and that of RSD during the day was 0.84 ~ 3.98, which was less than that of the commercial test kit of ELISA. Comparison of the results of BSA content in foods sold on the market, [conclusion] SPR method is reliable and stable, which is suitable for high throughput real-time detection of allergen BSA in food.
【作者单位】：南京农业大学国家肉品质量安全控制工程技术研究中心/农业部肉及肉制品质量监督检验测试中心(南京)/食品安全与营养协同创新中心;
【基金】：国家自然科学基金项目(31501395) 江苏省自然科学基金项目(BK20140722)
【分类号】：TP212;TS207.3

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