Jyoung Jy-Young, Hong SaHyun, Lee Woochang, Choi Jeong-Woo
Department of Chemical and Biomolecular Engineering, Sogang University, 1 Shinsu-Dong, Mapo-Gu, Seoul 121-742, Republic of Korea.
Biosens Bioelectron. 2006 Jun 15;21(12):2315-9. doi: 10.1016/j.bios.2005.10.015. Epub 2005 Dec 1.
An immunosensor for the detection of Vibrio cholerae O1 was developed on the basis of surface plasmon resonance (SPR). A protein G layer was fabricated by means of the chemical coupling between the free amine (-NH2) groups of protein G and the activated carboxyl groups present on a self-assembled monolayer (SAM) consisting of a mixture of 11-mercaptoundecanoic acid (MUA) and hexanethiol (molar ratio of 1:2). A monoclonal antibody, which was confirmed to be specific to V. cholera O1 by the Western blotting technique, was immobilized on the protein G layer. The formation of the SAM, the protein G layer and the sequential binding of the antibody against V. cholera O1 were investigated with SPR spectroscopy. As the number of fabricated layers increased, the minimum angle of plasmon resonance was increased accordingly. The target bacteria, V. cholera O1, was measured with the fabricated immunosensor, whose detection range was between 10(5) and 10(9) cells/mL.
基于表面等离子体共振（SPR）技术开发了一种用于检测霍乱弧菌O1的免疫传感器。通过蛋白质G的游离胺基（-NH2）与由11-巯基十一烷酸（MUA）和己硫醇（摩尔比1:2）混合物组成的自组装单分子层（SAM）上存在的活化羧基之间的化学偶联，制备了蛋白质G层。通过蛋白质印迹技术确认对霍乱弧菌O1具有特异性的单克隆抗体被固定在蛋白质G层上。利用SPR光谱研究了SAM、蛋白质G层的形成以及抗霍乱弧菌O1抗体的顺序结合。随着制备层数的增加，等离子体共振的最小角度相应增加。使用制备的免疫传感器对目标细菌霍乱弧菌O1进行了检测，其检测范围为10(5)至10(9)个细胞/mL。
