Rapid and label-free detection of protein a by aptamer-tethered porous silicon nanostructures

被引:38
|
作者
Urmann, Katharina [1 ,2 ]
Reich, Peggy [1 ,3 ]
Walter, Johanna-Gabriela [1 ]
Beckmann, Dieter [3 ]
Segal, Ester [2 ]
Scheper, Thomas [1 ]
机构
[1] Leibniz Univ Hannover, Inst Tech Chem, Callinstr 5, D-30167 Hannover, Germany
[2] Technion Israel Inst Technol, Dept Biotechnol & Food Engn, IL-32000 Haifa, Israel
[3] Inst Bioproc & Analyt Measurement Tech eV, D-37308 Rosenhof, Heilbad Heilige, Germany
基金
以色列科学基金会;
关键词
Optical biosensor; Aptamer; Porous silicon; Label-free; Protein A; Staphylococcus aureusa; FOURIER-TRANSFORM SPECTROSCOPY; STAPHYLOCOCCUS-AUREUS; OPTICAL BIOSENSOR; BINDING; SURFACE; IGG; IMMOBILIZATION; NANOPARTICLES; FLUORESCENCE; APTASENSOR;
D O I
10.1016/j.jbiotec.2017.01.005
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Protein A, which is secreted by and displayed on the cell membrane of Staphylococcus aureus is an important biomarker for S. aureus. Thus, its rapid and specific detection may facilitate the pathogen identification and initiation of proper treatment. Herein, we present a simple, label-free and rapid optical biosensor enabling specific detection of protein A. Protein A-binding aptamer serves as the capture probe and is immobilized onto a nanostructured porous silicon thin film, which serves as the optical transducer element. We demonstrate high sensitivity of the biosensor with a linear detection range between 8 and 23 mu M. The apparent dissociation constant was determined as 13.98 mu M and the LoD is 3.17 mu M. Harnessing the affinity between protein A and antibodies, a sandwich assay format was developed to amplify the optical signal associated with protein A capture by the aptamer. Using this approach, we increase the sensitivity of the biosensor, resulting in a three times lower LoD. (C) 2017 Elsevier B.V. All rights reserved.
引用
收藏
页码:171 / 177
页数:7
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