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

Hsueh Hsia Wu, Xin Quan Liao, Xin Ying Wu, Cheng De Lin, Kai Ru Jheng, Hong Ren Chen, Yong Yi Wang, Hsiu Mei Chen

研究成果: 雜誌貢獻文章

摘要

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.
原文英語
文章編號4493
期刊Sensors (Switzerland)
18
發行號12
DOIs
出版狀態已發佈 - 十二月 1 2018

指紋

Purple Membrane
Immunosensors
Graphite
Shear flow
Transducers
immobilization
shear flow
Immobilization
Oxides
Graphene
affinity
Monolayers
graphene
Staphylococcal Protein A
transducers
membranes
proteins
Proteins
Membranes
oxides

ASJC Scopus subject areas

  • Analytical Chemistry
  • Atomic and Molecular Physics, and Optics
  • Biochemistry
  • Instrumentation
  • Electrical and Electronic Engineering

引用此文

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. / Wu, Hsueh Hsia; Liao, Xin Quan; Wu, Xin Ying; Lin, Cheng De; Jheng, Kai Ru; Chen, Hong Ren; Wang, Yong Yi; Chen, Hsiu Mei.

於: Sensors (Switzerland), 卷 18, 編號 12, 4493, 01.12.2018.

研究成果: 雜誌貢獻文章

Wu, Hsueh Hsia ; Liao, Xin Quan ; Wu, Xin Ying ; Lin, Cheng De ; Jheng, Kai Ru ; Chen, Hong Ren ; Wang, Yong Yi ; Chen, Hsiu Mei. / 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. 於: Sensors (Switzerland). 2018 ; 卷 18, 編號 12.
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abstract = "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.",
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AU - Wu, Hsueh Hsia

AU - Liao, Xin Quan

AU - Wu, Xin Ying

AU - Lin, Cheng De

AU - Jheng, Kai Ru

AU - Chen, Hong Ren

AU - Wang, Yong Yi

AU - Chen, Hsiu Mei

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AB - 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.

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KW - Graphene oxide

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KW - Photoelectric immunosensor

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KW - Purple membrane

KW - Shear flow

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