Versatile Protein-A Coated Photoelectric Immunosensors with a Purple-Membrane Monolayer Transducer Fabricated by Affinity-Immobilization on a Graphene-Oxide Complexed Linker and by Shear Flow
| Autor: | Hsiu Mei Chen, Cheng De Lin, Hsueh Hsia Wu, Kai Ru Jheng, Xin Ying Wu, Yong Yi Wang, Xin Quan Liao, Hong Ren Chen |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
Materials science
shear flow Oxidized Avidin monolayer fabrication 02 engineering and technology photoelectric immunosensor protein A lcsh:Chemical technology 01 natural sciences Biochemistry Article Analytical Chemistry law.invention law Monolayer lcsh:TP1-1185 Electrical and Electronic Engineering Instrumentation Graphene bacteriorhodopsin 010401 analytical chemistry technology industry and agriculture Biological membrane 021001 nanoscience & nanotechnology Atomic and Molecular Physics and Optics 0104 chemical sciences Membrane Chemical engineering Biotinylation graphene oxide 0210 nano-technology Linker Biosensor purple membrane |
| Zdroj: | Sensors Volume 18 Issue 12 Sensors, Vol 18, Iss 12, p 4493 (2018) Sensors (Basel, Switzerland) |
| ISSN: | 1424-8220 |
| DOI: | 10.3390/s18124493 |
| Popis: | Bacteriorhodopsin-embedded purple membranes (PM) have been demonstrated to be a sensitive photoelectric transducer for microbial detection. To efficiently prepare versatile BR-based immunosensors with protein A as antibody captures, a large, high-coverage, and uniformly oriented PM monolayer was fabricated on an electrode as an effective foundation for protein A conjugation through bis-NHS esters, by first affinity-coating biotinylated PM on an aminated surface using a complex of oxidized avidin and graphene oxide as the planar linker and then washing the coating with a shear flow. Three different polyclonal antibodies, each against Escherichia coli, Lactobacillus acidophilus, and Streptococcus mutans, respectively, were individually, effectively and readily adsorbed on the protein A coated electrodes, leading to selective and sensitive quantitative detection of their respective target cells in a single step without any labeling. A single-cell detection limit was achieved for the former two cells. AFM, photocurrent, and Raman analyses all displayed each fabricated layer as well as the captured bacteria, with AFM particularly revealing the formation of a massive continuous PM monolayer on aminated mica. The facile cell-membrane monolayer fabrication and membrane surface conjugation techniques disclosed in this study may be widely applied to the preparation of different biomembrane-based biosensors. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
| Nepřihlášeným uživatelům se plný text nezobrazuje | K zobrazení výsledku je třeba se přihlásit. |