{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,7]],"date-time":"2026-06-07T01:55:14Z","timestamp":1780797314587,"version":"3.54.1"},"reference-count":27,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2022,2,24]],"date-time":"2022-02-24T00:00:00Z","timestamp":1645660800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"UNC\/NCSU Joint Department of Biomedical Engineering","award":["NA"],"award-info":[{"award-number":["NA"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>Electrochemical aptamer-based biosensors (E-ABs) are attractive candidates for use in biomarker detection systems due to their sensitivity, rapid response, and design flexibility. There are only several redox probes that were employed previously for this application, and a combination of redox probes affords some advantages in target detection. Thus, it would be advantageous to study new redox probes in an E-AB system. In this study, we report the use of amine-reactive phenazine ethosulfate (arPES) for E-AB through its conjugation to the terminus of thrombin-binding aptamer. The constructed E-AB can detect thrombin by square-wave voltammetry (SWV), showing peak current at \u22120.15 V vs. Ag\/AgCl at pH 7, which differs from redox probes used previously for E-ABs. We also compared the characteristics of PES as a redox probe for E-AB to methylene blue (MB), which is widely used. arPES showed stable signal at physiological pH. Moreover, the pH profile of arPES modified thrombin-binding aptamer revealed the potential application of arPES for a simultaneous multianalyte detection system. This could be achieved using different aptamers with several redox probes in tandem that harbor various electrochemical peak potentials. Our findings present a great opportunity to improve the current standard of biological fluid monitoring using E-AB.<\/jats:p>","DOI":"10.3390\/s22051760","type":"journal-article","created":{"date-parts":[[2022,2,24]],"date-time":"2022-02-24T21:11:07Z","timestamp":1645737067000},"page":"1760","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["An Amine-Reactive Phenazine Ethosulfate (arPES)\u2014A Novel Redox Probe for Electrochemical Aptamer-Based Sensor"],"prefix":"10.3390","volume":"22","author":[{"given":"Madoka","family":"Nagata","sequence":"first","affiliation":[{"name":"Joint Department of Biomedical Engineering, The University of North Carolina at Chapel Hill and North Carolina State University, Chapel Hill, NC 27599, USA"},{"name":"Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology, Koganei 184-8588, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinhee","family":"Lee","sequence":"additional","affiliation":[{"name":"Joint Department of Biomedical Engineering, The University of North Carolina at Chapel Hill and North Carolina State University, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen","family":"Henley","sequence":"additional","affiliation":[{"name":"Joint Department of Biomedical Engineering, The University of North Carolina at Chapel Hill and North Carolina State University, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2838-0562","authenticated-orcid":false,"given":"Kazunori","family":"Ikebukuro","sequence":"additional","affiliation":[{"name":"Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology, Koganei 184-8588, Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9833-2091","authenticated-orcid":false,"given":"Koji","family":"Sode","sequence":"additional","affiliation":[{"name":"Joint Department of Biomedical Engineering, The University of North Carolina at Chapel Hill and North Carolina State University, Chapel Hill, NC 27599, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,2,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1126\/science.2200121","article-title":"Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophageT4 DNA polymerase","volume":"249","author":"Tuerk","year":"1990","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"818","DOI":"10.1038\/346818a0","article-title":"In Vitro selection of RNA molecules that bind specific ligands","volume":"346","author":"Ellington","year":"1990","journal-title":"Nature"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1146\/annurev-anchem-071015-041446","article-title":"Reagentless, structure-switching, electrochemical aptamer-based sensors","volume":"9","author":"Macazo","year":"2016","journal-title":"Annu. Rev. Anal. Chem."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1021\/ar900165x","article-title":"Folding-based electrochemical biosensors: The case for responsive nucleic acid architectures","volume":"43","author":"Lubin","year":"2010","journal-title":"Acc. Chem. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5592","DOI":"10.1002\/ange.200500989","article-title":"Label-free electronic detection of thrombin in blood serum by using aptamer-based sensor","volume":"117","author":"Xiao","year":"2005","journal-title":"Angew. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3138","DOI":"10.1021\/ja056957p","article-title":"An electronic, aptamer-based small-molecule sensor for the rapid, label-free detection of cocaine in adulterated samples and biological fluids","volume":"128","author":"Baker","year":"2006","journal-title":"J. Am. Chem. Soc."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1021\/bc960070o","article-title":"Electrochemistry of methylene blue bound to a DNA-modified electrode","volume":"8","author":"Kelley","year":"1997","journal-title":"Bioconjugate Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1261","DOI":"10.1002\/elan.200804558","article-title":"Optimization of the electrochemical RNA-aptamer based biosensor for theophylline by using a methylene blue redox label","volume":"21","author":"Ferapontova","year":"2008","journal-title":"Electroanalysis"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1021\/ja053121d","article-title":"Reagentless, reusable, ultrasensitive electrochemical molecular beacon aptasensor","volume":"128","author":"Radi","year":"2006","journal-title":"J. Am. Chem. Soc."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1042","DOI":"10.1021\/ja067024b","article-title":"A Target-responsive electrochemical aptamer switch (TREAS) for reagentless detection of nanomolar ATP","volume":"129","author":"Zuo","year":"2007","journal-title":"J. Am. Chem. Soc."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.bios.2014.08.034","article-title":"Detecting multiple cell-secreted cytokines from the same aptamer-functionalized electrode","volume":"64","author":"Liu","year":"2014","journal-title":"Biosens. Bioelectron."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.bioelechem.2018.02.001","article-title":"Development of a glucose sensor employing quick and easy modification method with mediator for altering electron acceptor preference","volume":"121","author":"Hatada","year":"2018","journal-title":"Bioelectrochemistry"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"111974","DOI":"10.1016\/j.bios.2019.111974","article-title":"Rational engineering of Aerococcus viridans L-lactate oxidase for the mediator modification to achieve quasi-direct electron transfer type lactate sensor","volume":"151","author":"Hiraka","year":"2020","journal-title":"Biosens. Bioelectron."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Suzuki, N., Lee, J., Loew, N., Takahashi-Inose, Y., Okuda-Shimazaki, J., Kojima, K., Mori, K., Tsugawa, W., and Sode, K. (2020). Engineered glucose oxidase capable of quasi-direct electron transfer after a quick-and-easy modification with a mediator. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21031137"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"112984","DOI":"10.1016\/j.bios.2021.112984","article-title":"Development of glycated peptide enzyme sensor based flow injection analysis system for haemoglobin A1c monitoring using quasi-direct electron transfer type engineered fructosyl peptide oxidase","volume":"177","author":"Hatada","year":"2021","journal-title":"Biosens. Bioelectron."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"130554","DOI":"10.1016\/j.snb.2021.130554","article-title":"Continuous electrochemical monitoring of L-glutamine using redox-probe-modified L-glutamine-binding protein based on intermittent pulse amperometry","volume":"346","author":"Takamatsu","year":"2021","journal-title":"Sens. Actuators B. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"564","DOI":"10.1038\/355564a0","article-title":"Selection of single-stranded DNA molecules that bind and inhibit human thrombin","volume":"355","author":"Bock","year":"1992","journal-title":"Nature"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2875","DOI":"10.1038\/nprot.2007.413","article-title":"Preparation of electrode-immobilized, redox-modified oligonucleotides for electrochemical DNA and aptamer-based sensing","volume":"2","author":"Xiao","year":"2007","journal-title":"Nat. Protoc."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1021\/ac902595f","article-title":"Exploiting binding-induced changes in probe flexibility for the optimization of electrochemical biosensors","volume":"82","author":"White","year":"2010","journal-title":"Anal. Chem."},{"key":"ref_20","unstructured":"(2022, February 04). Dojindo, 1-Methoxy PES. Available online: https:\/\/dojindo.com\/product\/1-methoxy-pes-m470\/."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"113927","DOI":"10.1016\/j.bios.2021.113927","article-title":"Transient potentiometry based D-serine sensor using engineered D-amino acid oxidase showing quasi-direct electron transfer property","volume":"200","author":"Takamatsu","year":"2022","journal-title":"Biosens. Bioelectron."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.sbsr.2015.09.004","article-title":"Probing molecular interactions with methylene blue derivatized self-assembled monolayers","volume":"6","author":"Koutsoumpeli","year":"2015","journal-title":"Sens. Bio-Sens. Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1016\/j.bios.2016.10.089","article-title":"A novel \u201csignal-on\/off\u201d sensing platform for selective detection of thrombin based on target-induced ratiometric electrochemical biosensing and bio-bar-coded nanoprobe amplification strategy","volume":"92","author":"Wang","year":"2017","journal-title":"Biosens. Bioelectron."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4733","DOI":"10.1039\/c1cc10240k","article-title":"An aptamer-based signal-on and multiplexed sensing platform for one-spot simultaneous electronic detection of proteins and small molecules","volume":"47","author":"Xiang","year":"2011","journal-title":"Chem. Commun."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e1","DOI":"10.1038\/am.2012.1","article-title":"DNA biomolecular-electronic encoder and decoder devices constructed by multiplex biosensors","volume":"4","author":"Kang","year":"2012","journal-title":"NPG Asia Mater."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1016\/j.bios.2017.04.039","article-title":"Integrated signal probe based aptasensor for dual-analyte detection","volume":"96","author":"Xiang","year":"2017","journal-title":"Biosens. Bioelectron."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"15809","DOI":"10.1021\/jacs.6b08671","article-title":"Dual-reporter drift correction to enhance the performance of electrochemical aptamer-based sensors in whole blood","volume":"138","author":"Li","year":"2016","journal-title":"J. Am. Chem. Soc."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/22\/5\/1760\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T22:25:59Z","timestamp":1760135159000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/22\/5\/1760"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,2,24]]},"references-count":27,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2022,3]]}},"alternative-id":["s22051760"],"URL":"https:\/\/doi.org\/10.3390\/s22051760","relation":{},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,2,24]]}}}