Using poly(3-aminophenylboronic acid) thin film with binding-induced ion flux blocking for amperometric detection of hemoglobin A1c.

This study reports a novel enzyme-free, label-free amperometric method for direct detection of hemoglobin A1c (Hb(A1c)), a potent biomarker for diabetes diagnosis and prognosis. The method relies on an electrode modified with poly(3-aminophenylboronic acid) (PAPBA) nanoparticles (20-50 nm) and a sensing scheme named "binding-induced ion flux blocking." The PAPBA nanoparticles were characterized by FT-IR, XPS, TEM, and SEM. Being a polyaniline derivative, PAPBA showed an ion-dependent redox behavior, in which insertion or extraction of ions into or out of PABPA occurred for charge balance during the electron transfer process. The polymer allowed Hb(A1c) selectively bound to its surface via forming the cis-diol linkage between the boronic acid and sugar moieties. Voltammetric analyses showed that Hb(A1c) binding decreased the redox current of PAPBA; however, the binding did not alter the redox potentials and the apparent diffusivities of ions. This suggests that the redox current of PAPBA decreased due to an Hb(A1c) binding-induced ion flux blocking mechanism, which was then verified and characterized through an in situ electrochemical quartz crystal microbalance (EQCM) study. Assay with Hb(A1c) by differential pulse voltammetry (DPV) indicates that the peak current of a PAPBA electrode has a linear dependence on the logarithm of Hb(A1c) concentration ranging from 0.975 to 156 μM. The Hb(A1c) assay also showed high selectivity against ascorbic acid, dopamine, uric acid, glucose and bovine serum albumin. This study has demonstrated a new method for developing an electrochemical Hb(A1c) biosensor and can be extended to other label-free, indicator-free protein biosensors based on a similar redox polymer electrode.

[1]  J Wang,et al.  Processible polyaniline as an advanced potentiometric pH transducer. Application to biosensors. , 1999, Analytical chemistry.

[2]  D. Gjerde,et al.  Enrichment of Amadori products derived from the nonenzymatic glycation of proteins using microscale boronate affinity chromatography. , 2009, Analytical biochemistry.

[3]  Václav Kašička,et al.  Analysis of glycated hemoglobin A1c by capillary electrophoresis and capillary isoelectric focusing. , 2011, Analytical biochemistry.

[4]  L. Guariguata.  By the numbers: new estimates from the IDF Diabetes Atlas Update for 2012. , 2012, Diabetes research and clinical practice.

[5]  T. Wolever,et al.  Less variation of postprandial blood glucose after starchy test meals than oral glucose , 1996 .

[6]  Chandra Shekhar Pundir,et al.  Determination of glycated hemoglobin with special emphasis on biosensing methods. , 2014, Analytical biochemistry.

[7]  Tse-Chuan Chou,et al.  A Nonenzymatic Amperometric Method for Fructosyl‐Valine Sensing Using Ferroceneboronic Acid , 2011 .

[8]  M. Mohsen-Nia,et al.  Human Serum Protein Adsorption onto Synthesis Nano-Hydroxyapatite , 2012, The Protein Journal.

[9]  H. Bilo,et al.  Haemoglobin A1c: Historical overview and current concepts. , 2013, Diabetes research and clinical practice.

[10]  Yoon-Bo Shim,et al.  Disposable amperometric glycated hemoglobin sensor for the finger prick blood test. , 2013, Analytical chemistry.

[11]  Richard M Crooks,et al.  Determination of percent hemoglobin A1c using a potentiometric method. , 2013, Analytical chemistry.

[12]  Chandra Shekhar Pundir,et al.  An amperometric hemoglobin A1c biosensor based on immobilization of fructosyl amino acid oxidase onto zinc oxide nanoparticles-polypyrrole film. , 2012, Analytical biochemistry.

[13]  F. Arnold,et al.  The Temkin isotherm describes heterogeneous protein adsorption. , 1995, Biochimica et biophysica acta.

[14]  B. Sumerlin,et al.  Biomedical applications of boronic acid polymers , 2011 .

[15]  Asumi Tanaka,et al.  Relative kinetic reactivity of boronic acid and boronate ion towards Tiron, 2,2'-biphenol, and propylene glycol. , 2013, Dalton transactions.

[16]  P. Gillery.  A history of HbA1c through Clinical Chemistry and Laboratory Medicine , 2012, Clinical chemistry and laboratory medicine.

[17]  S. Tougaard,et al.  Comparison of validity and consistency of methods for quantitative XPS peak analysis , 1993 .

[18]  Ling-Sheng Jang,et al.  Detection of glycated hemoglobin (HbA1c) based on impedance measurement with parallel electrodes integrated into a microfluidic device , 2012 .

[19]  Tse-Chuan Chou,et al.  Amperometric protein sensor - fabricated as a polypyrrole, poly-aminophenylboronic acid bilayer. , 2006, Biosensors & bioelectronics.

[20]  Chandra Shekhar Pundir,et al.  An electrochemical biosensor for fructosyl valine for glycosylated hemoglobin detection based on core-shell magnetic bionanoparticles modified gold electrode. , 2011, Biosensors & bioelectronics.

[21]  Ming-Kun Chen,et al.  Glycated hemoglobin (HbA1c) affinity biosensors with ring-shaped interdigital electrodes on impedance measurement. , 2013, Biosensors & bioelectronics.

[22]  Frieder W. Scheller,et al.  Ferroceneboronic acid-based amperometric biosensor for glycated hemoglobin , 2006 .

[23]  A. C. Jamison,et al.  4-Mercaptophenylboronic acid SAMs on gold: comparison with SAMs derived from thiophenol, 4-mercaptophenol, and 4-mercaptobenzoic acid. , 2007, Langmuir : the ACS journal of surfaces and colloids.

[24]  E. Yeung,et al.  Separation of hemoglobin variants in single human erythrocytes by capillary electrophoresis with laser-induced native fluorescence detection. , 1995, Journal of chromatography. A.

[25]  M. Panteghini,et al.  Biological variability of glycated hemoglobin. , 2010, Clinica chimica acta; international journal of clinical chemistry.

[26]  Seung Yeon Song,et al.  Bioelectrocatalytic detection of glycated hemoglobin (HbA1c) based on the competitive binding of target and signaling glycoproteins to a boronate-modified surface. , 2012, Biosensors & bioelectronics.

[27]  Kuo-Chuan Ho,et al.  Synthesis of redox polymer nanobeads and nanocomposites for glucose biosensors. , 2013, ACS applied materials & interfaces.

[28]  Glucoregulation in type 2 diabetes: the lower the better? Glycosylated HbA1c of 6.5% seems to be the limit. , 2009, European journal of internal medicine.

[29]  S. K. Vashist.  Non-invasive glucose monitoring technology in diabetes management: a review. , 2012, Analytica chimica acta.

[30]  F. Frantzen,et al.  Protein-boronic acid conjugates and their binding to low-molecular-mass cis-diols and glycated hemoglobin. , 1995, Journal of chromatography. B, Biomedical applications.

[31]  Bo Mattiasson,et al.  Application of shielding boronate affinity chromatography in the study of the glycation pattern of haemoglobin. , 2002, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[32]  Frieder W. Scheller,et al.  Membrane-immobilized haptoglobin as affinity matrix for a hemoglobin-A1c immunosensor , 2002 .

[33]  Koji Sode,et al.  Development of a flow-injection analysis (FIA) enzyme sensor for fructosyl amine monitoring , 2002, Analytical and bioanalytical chemistry.

[34]  J. Jeppsson,et al.  Improved Method for Analysis of Glycated Haemoglobin by Ion Exchange Chromatography , 1994, Annals of clinical biochemistry.

[35]  Shanhong Xia,et al.  A micro-potentiometric hemoglobin immunosensor based on electropolymerized polypyrrole-gold nanoparticles composite. , 2009, Biosensors & bioelectronics.

[36]  Michael S Freund,et al.  Potentiometric saccharide detection based on the pK(a) changes of poly(aniline boronic acid). , 2002, Journal of the American Chemical Society.

[37]  M. Fahlman,et al.  Comparative XPS surface study of polyaniline thin films , 2008 .

[38]  Jing‐Juan Xu,et al.  Gold nanoparticles-coated magnetic microspheres as affinity matrix for detection of hemoglobin A1c in blood by microfluidic immunoassay. , 2011, Biosensors & bioelectronics.

[39]  J. Justin Gooding,et al.  An Electrochemical Impedance Immunosensor Based on Gold Nanoparticle‐Modified Electrodes for the Detection of HbA1c in Human Blood , 2012 .

[40]  Koji Sode,et al.  A new concept for the construction of an artificial dehydrogenase for fructosylamine compounds and its application for an amperometric fructosylamine sensor , 2001 .

[41]  Byoung-Yong Chang,et al.  Selective electrochemical sensing of glycated hemoglobin (HbA1c) on thiophene-3-boronic acid self-assembled monolayer covered gold electrodes. , 2008, Analytical chemistry.

[42]  S. L. Jeffcoate,et al.  Diabetes control and complications: the role of glycated haemoglobin, 25 years on , 2004, Diabetic medicine : a journal of the British Diabetic Association.

[43]  T. Fisher,et al.  Microwave‐Assisted Surface Synthesis of a Boron–Carbon–Nitrogen Foam and its Desorption Enthalpy , 2012 .