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通过局域表面等离子体光谱法测量抗体与固定在金纳米颗粒上的蛋白质的结合。

Measurement of antibody binding to protein immobilized on gold nanoparticles by localized surface plasmon spectroscopy.

作者信息

Fujiwara Kazuhiko, Watarai Hitoshi, Itoh Hideaki, Nakahama Erika, Ogawa Nobuaki

机构信息

Department of Materials Process & Applied Chemistry for Environments, Faculty of Engineering and Resource Science, Akita University, Tegata Gakuen-cho, Akita, 010-8502, Japan.

出版信息

Anal Bioanal Chem. 2006 Oct;386(3):639-44. doi: 10.1007/s00216-006-0559-2. Epub 2006 Jul 6.

Abstract

Antibody binding to bovine serum albumin (BSA) and human serum albumin (HSA) immobilized onto gold nanoparticles was studied by means of localized surface plasmon resonance (LSPR) spectroscopy. Amine-modified glass was prepared by self-assembly of amine-terminated silane on substrate, and gold (Au) nanoparticles were deposited on the amine-modified glass substrate. Au nanoparticles deposited on the glass surface were functionalized by BSA and HSA. BSA immobilization was confirmed by LSPR spectroscopy in conjunction with surface-enhanced Raman scattering spectroscopy. Then, LSPR response attributable to the binding of anti-BSA and anti-HSA to BSA- and HSA-functionalized Au nanoparticles, respectively, was examined. Anti-HSA at levels larger than approximately 10 nM could be detected by HSA-immobilized chips with LSPR optical response, which was saturated at concentrations greater than approximately 650 nM of anti-HSA.

摘要

通过局域表面等离子体共振(LSPR)光谱研究了抗体与固定在金纳米颗粒上的牛血清白蛋白(BSA)和人血清白蛋白(HSA)的结合情况。通过胺基封端的硅烷在基底上自组装制备胺基修饰的玻璃,然后将金(Au)纳米颗粒沉积在胺基修饰的玻璃基底上。沉积在玻璃表面的金纳米颗粒用BSA和HSA进行功能化修饰。结合表面增强拉曼散射光谱,通过LSPR光谱证实了BSA的固定化。然后,分别研究了抗BSA和抗HSA与BSA功能化和HSA功能化的金纳米颗粒结合所引起的LSPR响应。对于固定有HSA的芯片,当抗HSA浓度大于约10 nM时,可通过LSPR光学响应检测到抗HSA,当抗HSA浓度大于约650 nM时,LSPR光学响应达到饱和。

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