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用于检测鼠伤寒沙门氏菌的表面等离子体共振免疫传感器。

Surface plasmon resonance immunosensor for the detection of Salmonella typhimurium.

作者信息

Oh Byung-Keun, Kim Young-Kee, Park Kwang Won, Lee Won Hong, Choi Jeong-Woo

机构信息

Department of Chemical and Biomolecular Engineering, Sogang University, 1 Shinsu-dong, Mapo-gu, Seoul 121-742, South Korea.

出版信息

Biosens Bioelectron. 2004 Jun 15;19(11):1497-504. doi: 10.1016/j.bios.2003.12.009.

Abstract

An immunosensor based on surface plasmon resonance (SPR) using protein G was developed for the detection of Salmonella typhimurium. A protein G layer was fabricated by binding chemically to self-assembly monolayer (SAM) of 11-mercaptoundecanoic acid (MUA) on gold (Au) surface. The formation of protein G layer on Au surface modified with 11-MUA and the binding of antibody and antigen in series were confirmed by SPR spectroscopy. The effect of detergent such as Tween-20 on binding efficiency of antibody and antigen was investigated by SPR. The binding efficiency of antigen to the antibody immobilized on Au surface was improved up to about 85% and 100% by using protein G and Tween-20, respectively. The surface morphology analyses of 11-MUA monolayer on Au substrate, protein G layer on 11-MUA monolayer and antibody layer immobilized on protein G layer were performed by atomic force microscope (AFM). Consequently, an immunosensor based on SPR for the detection of S. typhimurium using protein G was developed with a detection range of 10(2) to 10(9)CFU/ml. The current fabrication technique of a SPR immunosensor for the detection of S. typhimurium could be applied to construct other immnosensors or protein chips.

摘要

开发了一种基于表面等离子体共振(SPR)并使用蛋白G的免疫传感器,用于检测鼠伤寒沙门氏菌。通过化学结合在金(Au)表面的11-巯基十一烷酸(MUA)自组装单分子层上制备蛋白G层。通过SPR光谱证实了在经11-MUA修饰的Au表面上蛋白G层的形成以及抗体和抗原的串联结合。通过SPR研究了吐温20等去污剂对抗体和抗原结合效率的影响。使用蛋白G和吐温20后,抗原与固定在Au表面的抗体的结合效率分别提高到约85%和100%。通过原子力显微镜(AFM)对Au基底上的11-MUA单分子层、11-MUA单分子层上的蛋白G层以及固定在蛋白G层上的抗体层进行了表面形态分析。因此,开发了一种基于SPR并使用蛋白G检测鼠伤寒沙门氏菌的免疫传感器,检测范围为10(2)至10(9)CFU/ml。目前用于检测鼠伤寒沙门氏菌的SPR免疫传感器的制造技术可应用于构建其他免疫传感器或蛋白质芯片。

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