{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T03:23:11Z","timestamp":1768447391054,"version":"3.49.0"},"reference-count":40,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2018,12,18]],"date-time":"2018-12-18T00:00:00Z","timestamp":1545091200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004663","name":"Ministry of Science and Technology, Taiwan","doi-asserted-by":"publisher","award":["MOST 105-2221-E-011-132"],"award-info":[{"award-number":["MOST 105-2221-E-011-132"]}],"id":[{"id":"10.13039\/501100004663","id-type":"DOI","asserted-by":"publisher"}]},{"name":"NTUST- TMU Joint Research Program","award":["NTUST-TMU-No. 107-04"],"award-info":[{"award-number":["NTUST-TMU-No. 107-04"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>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.<\/jats:p>","DOI":"10.3390\/s18124493","type":"journal-article","created":{"date-parts":[[2018,12,19]],"date-time":"2018-12-19T12:12:44Z","timestamp":1545221564000},"page":"4493","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["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"],"prefix":"10.3390","volume":"18","author":[{"given":"Hsueh-Hsia","family":"Wu","sequence":"first","affiliation":[{"name":"School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 11031, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xin-Quan","family":"Liao","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 10607, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xin-Ying","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 10607, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cheng-De","family":"Lin","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 10607, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kai-Ru","family":"Jheng","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 10607, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hong-Ren","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 10607, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yong-Yi","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 10607, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hsiu-Mei","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Chemical Engineering, National Taiwan University of Science and Technology, Taipei 10607, Taiwan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2018,12,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Zourob, M., Elwary, S., and Turner, A. (2008). Antibodies and immunoassays for detection of bacterial pathogens. Principles of Bacterial Detection: Biosensors, Recognition Receptors and Microsystems, Springer.","DOI":"10.1007\/978-0-387-75113-9"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"4407","DOI":"10.3390\/s90604407","article-title":"Antibody-based sensors: Principles, problems and potential for detection of pathogens and associated toxins","volume":"9","author":"Byrne","year":"2009","journal-title":"Sensors"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"631","DOI":"10.1128\/CMR.00120-13","article-title":"Biosensors for whole-cell bacterial detection","volume":"27","author":"Ahmed","year":"2014","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.ab.2009.05.029","article-title":"A biotin-hydrogel coated QCM biosensor and applications in immunoassay and peptide-displaying cell detection","volume":"392","author":"Chen","year":"2009","journal-title":"Anal. Biochem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1016\/j.bios.2013.01.009","article-title":"Highly sensitive detection of pathogen Escherichia coli O157:H7 by electrochemical impedance spectroscopy","volume":"45","author":"Agusil","year":"2013","journal-title":"Biosens. Bioelectron."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1016\/j.bios.2013.11.016","article-title":"Microconductometric immunosensor for label-free and sensitive detection of Gram-negative bacteria","volume":"54","author":"Leon","year":"2014","journal-title":"Biosens. Bioelectron."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.bios.2016.12.032","article-title":"Direct, label-free, selective, and sensitive microbial detection using a bacteriorhodopsin-based photoelectric immunosensor","volume":"91","author":"Chen","year":"2017","journal-title":"Biosens. Bioelectron."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1755","DOI":"10.1021\/cr980072x","article-title":"Bacteriorhodopsin as a photochromic retinal protein for optical memories","volume":"100","author":"Hampp","year":"2000","journal-title":"Chem. Rev."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"20130197","DOI":"10.1098\/rsif.2013.0197","article-title":"Directed evolution of bacteriorhodopsin for applications in bioelectronics","volume":"10","author":"Wagner","year":"2013","journal-title":"J. R. Soc. Interface"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.jtice.2016.03.040","article-title":"Intercalating purple membranes into 2D-alanine crystals to enhance photoelectric and nonlinear optical properties","volume":"64","author":"Chen","year":"2016","journal-title":"J. Taiwan Inst. Chem. Eng."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"822","DOI":"10.4049\/jimmunol.97.6.822","article-title":"\u201cProtein A\u201d from S. aureus. I. Pseudo-immune reaction with human-globulin","volume":"97","author":"Forsgren","year":"1966","journal-title":"J. Immunol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2769","DOI":"10.1039\/an9941902769","article-title":"Use of protein A as an immunological reagent and its application using flow injection. A review","volume":"119","author":"Palmer","year":"1994","journal-title":"Analyst"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1186\/s13036-015-0014-z","article-title":"Gold nanoparticle-labeled biosensor for rapid and sensitive detection of bacterial pathogens","volume":"9","author":"Wang","year":"2015","journal-title":"J. Biol. Eng."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1142\/S2339547816500072","article-title":"Immunosensor for rapid extraction\/detection of enteric pathogens","volume":"4","author":"Alocilja","year":"2016","journal-title":"Technology"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2642","DOI":"10.1039\/c3ay42036a","article-title":"Rapid biosensing of Staphylococcus aureus bacteria in milk","volume":"6","author":"Peedel","year":"2014","journal-title":"Anal. Methods"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"840","DOI":"10.1016\/j.bios.2005.01.021","article-title":"A QCM immunosensor for Salmonella detection with simultaneous measurements of resonant frequency and motional resistance","volume":"21","author":"Su","year":"2005","journal-title":"Biosen. Bioelectron."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1330","DOI":"10.1016\/j.bios.2008.07.071","article-title":"Rapid detection of Bacillus anthracis using monoclonal antibody functionalized QCM sensor","volume":"24","author":"Hao","year":"2009","journal-title":"Biosens. Bioelectron."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"79","DOI":"10.3390\/s150100079","article-title":"Rapid detection of microorganisms based on active and passive modes of QCM","volume":"15","author":"Farka","year":"2015","journal-title":"Sensors"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/S0003-2697(02)00442-6","article-title":"Optimizing antibody immobilization strategies for the construction of protein microarrays","volume":"312","author":"Peluso","year":"2003","journal-title":"Anal. Biochem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"e370","DOI":"10.4158\/EP161261.CR","article-title":"High-dose biotin treatment for secondary progressive multiple sclerosis nay interfere with thyroid assays","volume":"2","author":"Minkovsky","year":"2016","journal-title":"AACE Clin. Case Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3251","DOI":"10.1210\/jc.2016-1971","article-title":"Factitious Graves\u2019 disease due to biotin immunoassay interference\u2014A case and review of the literature","volume":"101","author":"Elston","year":"2016","journal-title":"J. Clin. Endocrinol. Metab."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"482","DOI":"10.1016\/j.colsurfb.2014.01.035","article-title":"Effect of graphene oxide on affinity-immobilization of purple membranes on solid supports","volume":"116","author":"Chen","year":"2014","journal-title":"Colloids Surf. B Biointerface"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3420","DOI":"10.1021\/jp962111j","article-title":"Molecular mechanism of the differential photoelectric response of bacteriorhodopsin","volume":"101","author":"Wang","year":"1997","journal-title":"J. Phys. Chem. B"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1695","DOI":"10.1016\/S0021-9258(17)43463-6","article-title":"Complete sequence of the staphylococcal gene encoding protein A. A gene evolved through multiple duplications","volume":"259","author":"Guss","year":"1984","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"338","DOI":"10.1016\/j.ijbiomac.2014.02.054","article-title":"Assessment of morphology and property of graphene oxide-hydroxypropylmethylcellulose nanocomposite films","volume":"66","author":"Ghosh","year":"2014","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"7416","DOI":"10.1021\/la900350s","article-title":"In situ IR spectroscopic studies of the avidin-biotin bioconjugation reaction on CdS particle films","volume":"25","author":"Young","year":"2009","journal-title":"Langmuir"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"9090","DOI":"10.1074\/jbc.M109.080457","article-title":"Biochemical and biological characterization of a new oxidized avidin with enhanced tissue binding properties","volume":"285","author":"Verdoliva","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"9855","DOI":"10.1039\/c2sm25938a","article-title":"Self-assembly of natural tripeptide glutathione triggered by graphene oxide","volume":"8","author":"Yang","year":"2012","journal-title":"Soft Matter"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2277","DOI":"10.1039\/b920539j","article-title":"The chemistry of graphene","volume":"20","author":"Loh","year":"2010","journal-title":"J. Mater. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"4641","DOI":"10.1039\/c0cp02143a","article-title":"On the nature of OH-stretching vibrations in hydrogen-bonded chains: Pump frequency dependent vibrational lifetime","volume":"13","author":"Knop","year":"2011","journal-title":"Phys. Chem. Chem. Phys."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1073","DOI":"10.1016\/j.bbabio.2007.06.004","article-title":"Infrared spectroscopy of proteins","volume":"1767","author":"Barth","year":"2007","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3166","DOI":"10.1021\/la00035a069","article-title":"Cooperativity in the binding of avidin to biotin-lipid-doped Langmuir-Blodgett films","volume":"9","author":"Zhao","year":"1993","journal-title":"Langmuir"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2075","DOI":"10.1529\/biophysj.105.072405","article-title":"Differential stiffness and lipid mobility in the leaflets of purple membranes","volume":"90","author":"Contera","year":"2006","journal-title":"Biophys. J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1021\/nl071822y","article-title":"Raman spectra of graphite oxide and functionalized graphene sheets","volume":"8","author":"Kudin","year":"2008","journal-title":"Nano Lett."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1016\/S0006-3495(98)77803-8","article-title":"Correlation between surface morphology and surface forces of protein A adsorbed on mica","volume":"74","author":"Ohnishi","year":"1998","journal-title":"Biophys. J."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/S0927-7765(98)00055-1","article-title":"Binding of immunoglobulin molecules to preadsorbed protein A layers as observed by surface forces measurements","volume":"12","author":"Murata","year":"1998","journal-title":"Colloids Surf. B Biointerface"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/j.jphotobiol.2008.05.009","article-title":"Asymmetric distribution of biotin labeling on the purple membrane","volume":"92","author":"Su","year":"2008","journal-title":"J. Photochem. Photobiol. B"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1702","DOI":"10.1126\/science.1067172","article-title":"Protein nanoarrays generated by dip-pen nanolithography","volume":"295","author":"Lee","year":"2002","journal-title":"Science"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1016\/j.bios.2015.09.030","article-title":"Rapid detection of multiple foodborne pathogens using a nanoparticle-functionalized multi-junction biosensor","volume":"77","author":"Yamada","year":"2016","journal-title":"Biosens. Bioelectron."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0079-6816(99)00014-3","article-title":"Interfacial photochemistry of retinal proteins","volume":"62","author":"Hong","year":"1999","journal-title":"Prog. Surf. Sci."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/18\/12\/4493\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:34:49Z","timestamp":1760196889000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/18\/12\/4493"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,12,18]]},"references-count":40,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2018,12]]}},"alternative-id":["s18124493"],"URL":"https:\/\/doi.org\/10.3390\/s18124493","relation":{},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,12,18]]}}}