A Novel Label-Free Immunosensor Based on Electrochemically Reduced Graphene Oxide for Determination of Hemoglobin A 1c
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ovel Label-Free Immunosensor Based on Electrochemically Reduced Graphene Oxide for Determination of Hemoglobin A1c M. Özge Karaşallıa and Derya Koyuncu Zeybeka, * aKütahya
Dumlupınar University, Faculty of Science and Arts, Department of Biochemistry, Kütahya, Turkey *e-mail: [email protected] Received August 4, 2019; revised October 31, 2019; accepted December 20, 2019
Abstract—In this work, a novel immunosensor has been developed using an electrochemically reduced graphene oxide (ERGO) modified glassy carbon electrode (GCE) for sensitive and label-free detection of glycated hemoglobin (HbA1c) used as the gold standard marker for glycemic control. The prepared immunosensor, using ferrocyanide/ferricyanide as the mediator, ensures an uncomplicated and economic method for the detection of HbA1c. The proposed immunosensor can directly record changes in the electrochemical signals when the immunoreaction occurs on the sensor surface. The specific antibody-antigen immunoreaction resulted in a reduction in the amperometric response of the mediator that diffused at the electrode surface. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) techniques were utilized to investigate the electrochemical characteristics of the immunosensor. Calibration curve was plotted via differential pulse voltammetry (DPV) technique and a linear relationship was obtained between the DPV response and HbA1c concentrations from 1 to 25%. Under the optimized condition, the GCE/ERGO/HbA1cAb immunosensor supplied high selectivity and sensitivity for HbA1c detection in real serum samples. Keywords: label-free detection, electrochemical immunosensor, diabetes mellitus, graphene oxide, differential pulse voltammetry, HbA1c DOI: 10.1134/S1023193520090037
INTRODUCTION Diabetes Mellitus (DM) is characterized by chronic hyperglycemia affecting approximately 150 million people worldwide. According to data from the International Diabetes Federation, the number of people suffering with diabetes is anticipated to rise from 371 million in 2012 to 552 million by 2030. Chronic hyperglycemia can affect different organs, especially the heart, nerves, eyes, kidneys, and blood vessels. Thus, it is necessary to determine of the blood glucose level (BGL) for diagnosis and treatment of DM. The quantification of the fasting blood glucose and 2-h BGL is the most frequently utilized method for the diagnosis of DM. Nevertheless, when the BGL is detected in this way, its value may fluctuate, and be influenced by the daily diet, and should be quantified at organized intervals [1, 2]. Hemoglobin A1c (HbA1c) or glycated hemoglobin biomolecule is described as the gold standard for the diagnosis and monitoring of DM [3]. The HbA1c level represents the average BGL for 2‒3 months and is not influenced by daily fluctuations in the BGL. Consequently, the level of HbA1c is utilized for the diagnosis of diabetes glycemic control. Several conventional methods have been informed to
detection of HbA1c, such as boronate-affinity chromatography [4], capillar
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