{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T09:10:10Z","timestamp":1780477810816,"version":"3.54.1"},"reference-count":28,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2016,7,1]],"date-time":"2016-07-01T00:00:00Z","timestamp":1467331200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>A single-use disposable in vitro electrochemical immunosensor for the detection of HbA1c in undiluted human serum using differential pulse voltammetry (DPV) was developed. A three-electrode configuration electrochemical biosensor consisted of 10-nm-thin gold film working and counter electrodes and a thick-film printed Ag\/AgCl reference electrode was fabricated on a polyethylene terephthalate (PET) substrate. Micro-fabrication techniques including sputtering vapor deposition and thick-film printing were used to fabricate the biosensor. This was a roll-to-roll cost-effective manufacturing process making the single-use disposable in vitro HbA1c biosensor a reality. Self-assembled monolayers of 3-Mercaptopropionic acid (MPA) were employed to covalently immobilize anti-HbA1c on the surface of gold electrodes. Electrochemical impedance spectroscopy (EIS) and X-ray photoelectron spectroscopy (XPS) confirmed the excellent coverage of MPA-SAM and the upward orientation of carboxylic groups. The hindering effect of HbA1c on the ferricyanide\/ferrocyanide electron transfer reaction was exploited as the HbA1c detection mechanism. The biosensor showed a linear range of 7.5\u201320 \u00b5g\/mL of HbA1c in 0.1 M PBS. Using undiluted human serum as the test medium, the biosensor presented an excellent linear behavior (R2 = 0.999) in the range of 0.1\u20130.25 mg\/mL of HbA1c. The potential application of this biosensor for in vitro measurement of HbA1c for diabetic management was demonstrated.<\/jats:p>","DOI":"10.3390\/s16071024","type":"journal-article","created":{"date-parts":[[2016,7,1]],"date-time":"2016-07-01T09:51:48Z","timestamp":1467366708000},"page":"1024","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":60,"title":["Single-Use Disposable Electrochemical Label-Free Immunosensor for Detection of Glycated Hemoglobin (HbA1c) Using Differential Pulse Voltammetry (DPV)"],"prefix":"10.3390","volume":"16","author":[{"given":"Alireza","family":"Molazemhosseini","sequence":"first","affiliation":[{"name":"Dipartimento Chimica Materiali e Ingegneria Chimica \u201cGiulio Natta\u201d, Politecnico di Milano, Via Mancinelli 7, 20131 Milan, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luca","family":"Magagnin","sequence":"additional","affiliation":[{"name":"Dipartimento Chimica Materiali e Ingegneria Chimica \u201cGiulio Natta\u201d, Politecnico di Milano, Via Mancinelli 7, 20131 Milan, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pasquale","family":"Vena","sequence":"additional","affiliation":[{"name":"Dipartimento Chimica Materiali e Ingegneria Chimica \u201cGiulio Natta\u201d, Politecnico di Milano, Via Mancinelli 7, 20131 Milan, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chung-Chiun","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Chemical &amp; Biomolecular Engineering and Electronics Design Center, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, OH 44106, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2016,7,1]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1761","DOI":"10.2337\/diacare.27.7.1761","article-title":"Tests of glycemia in diabetes","volume":"27","author":"Goldstein","year":"2004","journal-title":"Diabetes Care"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"344","DOI":"10.1136\/jcp.2003.010918","article-title":"HbA1c measurement","volume":"57","author":"Kilpatrick","year":"2004","journal-title":"J. Clin. Pathol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/00365510500390130","article-title":"Haemoglobin A1c reference method","volume":"66","author":"John","year":"2006","journal-title":"Scand. J. Clin. Lab. Investig."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1199","DOI":"10.1515\/CCLM.2003.184","article-title":"Haemoglobin A1c: Analysis and standardisation","volume":"41","author":"John","year":"2003","journal-title":"Clin. Chem. Lab. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1515\/CCLM.2002.016","article-title":"Approved IFCC Reference Method for the Measurement of HbA1c in Human Blood","volume":"40","author":"Jeppsson","year":"2002","journal-title":"Clin. Chem. Lab. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1177\/000456329403100409","article-title":"Improved method for analysis of glycated haemoglobin by ion exchange chromatography","volume":"31","author":"Eckerbom","year":"1994","journal-title":"Ann. Clin. Biochem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2390","DOI":"10.1093\/clinchem\/43.12.2390","article-title":"Glycohemoglobin filter assay for doctors\u2019 offices based on boronic acid affinity principle","volume":"43","author":"Frantzen","year":"1997","journal-title":"Clin. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.cca.2015.03.025","article-title":"Evaluation of hemoglobin A1c measurement by Capillarys 2 electrophoresis for detection of abnormal glucose tolerance in African immigrants to the United States","volume":"446","author":"Zhao","year":"2015","journal-title":"Clin. Chim. Acta"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1016\/S0960-894X(03)00086-6","article-title":"A novel type of fluorescent boronic acid that shows large fluorescence intensity changes upon binding with a carbohydrate in aqueous solution at physiological pH","volume":"13","author":"Yang","year":"2003","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1145","DOI":"10.1093\/oxfordjournals.jbchem.a124835","article-title":"Novel Sensing System for Glucose Based on the Complex Formation between Phenylborate and Fluorescent Diol Compounds","volume":"117","author":"Kataoka","year":"1995","journal-title":"J. Biochem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1187","DOI":"10.3390\/ma8031187","article-title":"Recent Progress in Electrochemical HbA1c Sensors: A Review","volume":"8","author":"Wang","year":"2015","journal-title":"Materials"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1016\/j.bios.2014.09.058","article-title":"Fabrication of electrochemical interface based on boronic acid-modified pyrroloquinoline quinine\/reduced graphene oxide composites for voltammetric determination of glycated hemoglobin","volume":"64","author":"Zhou","year":"2014","journal-title":"Biosens. Bioelectron."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1002\/elan.201300646","article-title":"Point-of-Care Amperometric Testing of Diabetic Marker (HbA1c) Using Specific Electroactive Antibodies","volume":"26","author":"Chopra","year":"2014","journal-title":"Electroanalysis"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"6536","DOI":"10.1021\/ac401411y","article-title":"Disposable amperometric glycated hemoglobin sensor for the finger prick blood test","volume":"85","author":"Kim","year":"2013","journal-title":"Anal. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"8035","DOI":"10.1021\/ac8010439","article-title":"Selective Electrochemical Sensing of Glycated Hemoglobin (HbA(1c)) on Thiophene-3-Boronic Acid Self-Assembled Monolayer Covered Gold Electrodes","volume":"80","author":"Park","year":"2008","journal-title":"Anal. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1127","DOI":"10.1016\/j.electacta.2007.03.059","article-title":"Development of a biosensor for glycated hemoglobin","volume":"53","author":"Wollenberger","year":"2007","journal-title":"Electrochim. Acta"},{"key":"ref_17","unstructured":"Bhat, N., and Srinivasan, S. (2011, January 7\u20138). Detection of GlycatedHemoglobin using 3-AminoPhenylboronic acid modified Graphene Oxide. Proceedings of the IEEE\/NIH Life Science Systems and Applications Workshop (LiSSA), Bethesda, MD, USA."},{"key":"ref_18","first-page":"93","article-title":"Detection of Lysyl Oxidase-Like 2 (LOXL2), a Biomarker of Metastasis from Breast Cancers Using Human Blood Samples","volume":"5","author":"Janyasupab","year":"2015","journal-title":"Curr. Biomark."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1535","DOI":"10.1021\/ac00081a028","article-title":"Electrical Communication between Electrodes and NAD(P)+-Dependent Enzymes Using Pyrroloquinolinequinone-Enzyme Electrodes in a Self-Assembled Monolayer Configuration: Design of a New Class of Amperometric Biosensors","volume":"66","author":"Willner","year":"1994","journal-title":"Anal. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"92","DOI":"10.1016\/S0022-0728(02)00753-2","article-title":"Preparation, characterization and application of alkanethiol self-assembled monolayers modified with tetrathiafulvalene and glucose oxidase at a gold disk electrode","volume":"526","author":"Campuzano","year":"2002","journal-title":"J. Electroanal. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"940","DOI":"10.1039\/c2lc41049d","article-title":"Rapid and specific electrochemical detection of prostate cancer cells using an aperture sensor array","volume":"13","author":"Moscovici","year":"2013","journal-title":"Lab Chip"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/S0302-4598(96)05173-2","article-title":"Characterising the formation of a bioelectrochemical interface at a self-assembled monolayer using X-ray photoelectron spectroscopy","volume":"42","author":"Jiang","year":"1997","journal-title":"Bioelectrochem. Bioenerg."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.bioelechem.2008.12.005","article-title":"3-Mercaptopropylphosphonic Acid Modified Gold Electrode for Electrochemical Detection of Dopamine","volume":"75","author":"Chen","year":"2009","journal-title":"Bioelectrochemistry"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"6562","DOI":"10.1021\/jp9935337","article-title":"Gold-sulfur bonding in 2D and 3D self-assembled monolayers: XPS characterization","volume":"104","author":"Bourg","year":"2000","journal-title":"J. Phys. Chem. B"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.elspec.2009.12.007","article-title":"XAS and XPS examination of the Au-S nanostructures produced via the reduction of aqueous gold(III) by sulfide ions","volume":"177","author":"Mikhlin","year":"2010","journal-title":"J. Electron Spectrosc. Relat. Phenom."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"8830","DOI":"10.1021\/ac101474k","article-title":"Ternary surface monolayers for ultrasensitive (Zeptomole) amperometric detection of nucleic acid hybridization without signal amplification","volume":"82","author":"Wu","year":"2010","journal-title":"Anal. Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3011","DOI":"10.1021\/la970980+","article-title":"Impedance Spectroscopy of Self-Assembled Monolayers on Au(111): Sodium Ferrocyanide Charge Transfer at Modified Electrodes","volume":"14","author":"Janek","year":"1998","journal-title":"Langmuir"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1016\/j.snb.2009.04.057","article-title":"Boronic acid-modified thin film interface for specific binding of glycated hemoglobin (HbA1c) and electrochemical biosensing","volume":"140","author":"Song","year":"2009","journal-title":"Sens. Actuators B Chem."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/16\/7\/1024\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T19:25:18Z","timestamp":1760210718000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/16\/7\/1024"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,7,1]]},"references-count":28,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2016,7]]}},"alternative-id":["s16071024"],"URL":"https:\/\/doi.org\/10.3390\/s16071024","relation":{},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,7,1]]}}}