生物素-亲和素增敏抑制型SPR等离子共振传感器检测牛奶中链霉素  

Detection of Streptomycin in Milk by Biotin-Avidin Sensitization SPR Plasmon Resonance Sensor

作　　者：王敏思 柳双 朱文博 宋洋[1] Wang Minsi;Liu Shuang;Zhu Wenbo;Song Yang(Tianjin Key Laboratory of Animal and Plant Resistance,Tianjin Normal University,Tianjin 300387)

机构地区：[1]天津师范大学天津市动植物抗性重点实验室,天津300387

出　　处：《中国食品学报》2021年第10期262-268,共7页Journal of Chinese Institute Of Food Science and Technology

基　　金：天津市自然科学基金项目(18JCQNJC84400)。

摘　　要：针对牛奶中链霉素(STR)残留问题,开发一种基于生物素-亲和素系统(BAS)增敏及抑制型等离子共振(SPR)免疫传感器。方法:直接交联法获得的半抗原STR-OVA,通过活化酯法偶联生物素得到抗原BNHS-STR-OVA,与芯片上的链霉亲和素(SA)结合,绑定抗原。依据免疫抑制原理,STR与BNHS-STR-OVA共同竞争固定浓度的特异性抗体,依据不同浓度STR所得响应值,计算抑制率并绘制曲线,实现对食品中STR残留的准确、灵敏、快速分析。利用高效液相色谱串联质谱(HPLC-MS)检测该方法的准确性。结果:检测过程中抗原的固定量为6μg,抗体质量浓度为60 mg/mL。构建的抑制型免疫传感器的检出限(IC15)为0.37μg/L,灵敏度(IC50)为1.65μg/L。检测时长不超过10 min,芯片可重复使用90次。结论:研究所构建的BAS-SPR增敏、抑制型免疫传感器,方法简单,灵敏度高,检测时间短,成本低,可满足牛奶样品中STR高效检测的需求。This study has developed a sensitive and inhibitory surface plasmon resonance(SPR) immunosensor based on the Biotin-avidin system(BAS) for the detection of streptomycin(STR) residue in milk. Methods: STR hapten(STR-OVA) is prepared by direct cross-linking method, and coupled with biotin(BHNS) by activated ester method to obtain antigen(BNHS-STR-OVA), the antigen is combined with streptavidin(SA) on the SA chip. Based on the principle of immune suppression, the antibody for STR at a stable concentration was simultaneously bound by the STR and the antigen BNHS-STR-OVA. According to the SPR response value obtained under different STR concentretions, the inhibition ratio was calculated and the accurete, sensitivity and celerity analysis of STR in milk could be realized according to the inhibiting ratio curve. HPLC-MS was used to test the accuracy of the method. Results: the antigen was fixed at 6 μg;the mass concentration of antibody was established at 60 mg/mL;the detection limit(IC15) and the sensitivity(IC50) for STR using the proposed SPR immunosensor achieved to 0.37 μg/L and 1.65 μg/L;the detection process could be finished within 10 min, and one chip could be reused more than 90 times. Conclusion: BAS-SPR enhanced and suppressed SPR immunosensor has offered a simple, fast, low-cost detection strategy with high sensitivity for STR, which can meet the requirements of efficient detection of STR in milk samples.

关 键 词：链霉素 表面等离子体共振 生物素-亲和素 牛奶 

分 类 号：TS252.7[轻工技术与工程—农产品加工及贮藏工程] TP212[轻工技术与工程—食品科学与工程]