基于量子点的新型均相荧光免疫分析技术  

A novel quantum-dot-based homogeneous fluoroimmunoassay technology

作　　者：陈玫君[1] 陈振华[1] 吴英松[1] 任志奇[1] 李思雯[1] 刘天才[1] 

机构地区：[1]南方医科大学生物技术学院,广州510515

出　　处：《中国科技论文》2013年第6期487-491,共5页China Sciencepaper

基　　金：国家自然科学基金资助项目(30901382;81271931);高等学校博士学科点专项科研基金资助项目(20094433120008);广东省高等学校人才引进专项资金资助项目(2009);南方医科大学引进人才科研启动基金资助项目(2009)

摘　　要：研究一种检测人血清中癌胚抗原(CEA)的新型量子点均相免疫分析方法。制备还原型谷胱甘肽包被的水溶性量子点(QDs),偶联上抗CEA单克隆抗体,使异硫氰基荧光素(FITC)偶联上另一株抗CEA单克隆抗体,采用双抗体夹心法模式,充分利用两株抗体与CEA抗原的相互作用,从而验证QDs、FTTC两者与抗体偶联的生物活性,并研究QDs用于均相荧光共振能量转移(HFRET)免疫分析中的可行性与实用性。结果表明,FITC与QDs之间能够发生能量转移,其荧光共振能量转移信号与CEA浓度成线性相关,其工作曲线为LogY=2.504+0.154LogX(r2=0.981 97,P=0.002),并计算出其分析灵敏度为0.4ng/mL。该新型均相荧光免疫分析方法具有不需冲洗掉游离荧光标记物、灵敏度高等优势,为今后实现快速便捷、多通道、高灵敏度、微量化、自动化及高通量化的生物医学传感技术和现代免疫分析技术提供科学依据。A novel quantum-dot-based homogeneous fluoroimmunoassay technology has been studied for detection of carcinoem- bryonic antigen （CEA） in human serum. Water-soluble quantum dots （QDs） were prepared and coated with anti-CEA monoclonal antibody （McAb）, and another anti-CEA McAb was labeled with Fluorescein isothiocyanate （FITC）. The activities of these la- beled antibodies were analyzed by fluorescence resonance energy transfer （FRET）. The concentrations of CEA were measured by FRET between FITC and QDs with the two-antibody sandwich system. The results indicated that quantum dots could be applied in homogeneous fluoroimmunoassay. The signals of FRET were proportional to the concentrations of CEA with LogY = 2. 504＋ 0. 154LogX （r^2 = 0. 981 97, P = 0. 002）; the calculated sensitivity was 0. 4 ng/mL. The novel homogeneous fluorescence immunoassay does not require to wash away the free fluorescent conjugates with high sensitivity. It provides a scientific basis for the development of rapid and convenient biomedical sensor technology and modern immunoassay technology with multi-channel, high sensitivity and high-throughput.

关 键 词：量子点 双抗体夹心法 荧光共振能量转移 均相荧光免疫分析 

分 类 号：R392.1[医药卫生—免疫学]