摘要
建立一种基于间接竞争法的表面等离子共振(surface plasmon resonance,SPR)免疫传感技术检测雌二醇(estradiol,E_2),这种方法无需标记,可以实时监测竞争过程。在间接竞争实验中,E_2和E_2的单克隆抗体(monoclonal antibody,E_2-m Ab)等体积混合后共同竞争被固定在了SPR传感器的芯片表面的E_2-牛血清白蛋白包被原(E_2-bovine serum albumin,E_2-BSA),SPR产生的信号和E_2的质量浓度呈反比。优化实验参数如抗体质量浓度、再生次数等,最后再用全脂牛奶做E_2的加标实验。实验的线性范围为15.63~500 ng/m L,最低检出限为11.13 ng/m L。本研究的SPR免疫传感技术构建了一种通用的方法,可以用来无标记检测其他不同的小分子。
A surface plasmon resonance(SPR)biosensor system was developed for the determination of estradiol(E2)using an indirect competition method without labeling in real time.In the indirect competition method,the antigen(E2-BSA)was immobilized on the sensor chip surface and competed with E2in samples for binding with the antibodies.Thenthe SPR response decreased in the presence of E2,which prevented the coupling of antibodies and E2-BSA,and so the SPRresponse was inversely proportional to E2concentration.Assay parameters,including antibody concentration and number ofregenerations,were optimized and E2-spiked milk samples were detected by this method.A linear relationship was obtainedbetween inhibition percentage and common logarithmic value of E2concentration over the range of15.63to500ng/mL andthe limit of detection(LOD)was11.13ng/mL.The present study demonstrated a simple and general strategy for label-freedetection of various small molecules.
作者
贾映彤
彭媛
白家磊
张希浩
宁保安
高志贤
崔建升
JIA Yingtong;PENG Yuan;BAI Jialei;ZHANG Xihao;NING Bao-an;GAO Zhixian;CUI Jiansheng(School of Environmental Science and Engineering, Hebei University of Science and Technology, Shijiazhuang 050018, China;Tianjin Key Laboratory of Risk Assessment and Control Technology for Environment and Food Safety,Institute of Health and Environment Medicine, Academy of Military Medical Sciences, Tianjin 300050, China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2017年第12期223-228,共6页
Food Science
基金
国家自然科学基金青年科学基金项目(21207161)
国家自然科学基金面上项目(81472985)
军队重大专项(AWS15J006)
天津市应用基础与前沿技术研究计划项目(15JCYBJC51200)
关键词
表面等离子共振
无标记
雌二醇
实时
间接竞争
surface plasmon resonance (SPR)
label-free
estradiol
real-time
indirect competition
