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

Development of a surface plasmon resonance sensor for detection of animal allergen bovine serum albumin

[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(P>0.05)。[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。

[Objectives]A rapid,accurate and sensitive method for the quantitative detection of animal allergen bovine serum albumin（ BSA） was developed using surface plasmon resonance（ SPR） technology. [Methods]SPR was developed by monitoring the interaction between BSA and a self-prepared mouse anti-BSA monoclonal antibody on the SPR sensor chip,then determined for limit of detection（ LOD） and optimal linear range,verified for reproducibility and applied preliminarily. [Results]The specific monoclonal antibody against BSA at a labeling level of 18 000 showed good affinity to BSA. The BSA concentration at a range of 6. 25-400ng·m L^-1showed good linear relationship to the response value of SPR analyzer（ R^2= 0.992）,with LOD of 33.68 ng·m L^-1. The RSD of intra-and inter-assays on BSA at various concentrations by the developed SPR method was 1.12%-3.42% and 0.84%-3.98%,respectively,both of which were less than 5%. No significant differences were observed between the determination results of BSA of commercial meat and meat products by SPR and ELISA methods（ P〉0.05）. [Conclusions]The SPR method constructed in the paper has high reliability and stability,and can be used for high-throughput and real-time detection of animal allergen BSA in food.