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Direct and Label-Free Determination of Human Glycated Hemoglobin Levels Using Bacteriorhodopsin as the Biosensor Transducer

Glycated hemoglobin (HbA1c) levels are an important index for the diagnosis and long-term control of diabetes. This study is the first to use a direct and label-free photoelectric biosensor to determine HbA1c using bacteriorhodopsin-embedded purple membranes (PM) as a transducer. A biotinylated PM (...

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Autores principales: Lin, Ying-Chin, Lin, Ching-Yu, Chen, Hsiu-Mei, Kuo, Li-Pin, Hsieh, Cheng-En, Wang, Xiang-He, Cheng, Chih-Wen, Wu, Chih-Yin, Chen, Yi-Su
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7765918/
https://www.ncbi.nlm.nih.gov/pubmed/33353006
http://dx.doi.org/10.3390/s20247274
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author Lin, Ying-Chin
Lin, Ching-Yu
Chen, Hsiu-Mei
Kuo, Li-Pin
Hsieh, Cheng-En
Wang, Xiang-He
Cheng, Chih-Wen
Wu, Chih-Yin
Chen, Yi-Su
author_facet Lin, Ying-Chin
Lin, Ching-Yu
Chen, Hsiu-Mei
Kuo, Li-Pin
Hsieh, Cheng-En
Wang, Xiang-He
Cheng, Chih-Wen
Wu, Chih-Yin
Chen, Yi-Su
author_sort Lin, Ying-Chin
collection PubMed
description Glycated hemoglobin (HbA1c) levels are an important index for the diagnosis and long-term control of diabetes. This study is the first to use a direct and label-free photoelectric biosensor to determine HbA1c using bacteriorhodopsin-embedded purple membranes (PM) as a transducer. A biotinylated PM (b-PM) coated electrode that is layered with protein A-oriented antibodies against hemoglobin (Hb) readily captures non-glycated Hb (HbA0) and generates less photocurrent. The spectra of bacteriorhodopsin and Hb overlap so the photocurrent is reduced because of the partial absorption of the incident light by the captured Hb molecules. Two HbA0 and HbA1c aptasensors that are prepared by conjugating specific aptamers on b-PM coated electrodes single-step detect HbA0 and HbA1c in 15 min, without cross reactivity, with detection limits of ≤0.1 μg/mL and a dynamic range of 0.1–100 μg/mL. Both aptasensors exhibit high selectivity and long-term stability. For the clinical samples, HbA0 concentrations and HbA1c levels that are measured with aptasensors correlate well with total Hb concentrations and the HbA1c levels that are determined using standard methods (correlation gradient = 0.915 ± 0.004 and 0.981 ± 0.001, respectively). The use of these aptasensors for diabetes care is demonstrated.
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spelling pubmed-77659182020-12-28 Direct and Label-Free Determination of Human Glycated Hemoglobin Levels Using Bacteriorhodopsin as the Biosensor Transducer Lin, Ying-Chin Lin, Ching-Yu Chen, Hsiu-Mei Kuo, Li-Pin Hsieh, Cheng-En Wang, Xiang-He Cheng, Chih-Wen Wu, Chih-Yin Chen, Yi-Su Sensors (Basel) Article Glycated hemoglobin (HbA1c) levels are an important index for the diagnosis and long-term control of diabetes. This study is the first to use a direct and label-free photoelectric biosensor to determine HbA1c using bacteriorhodopsin-embedded purple membranes (PM) as a transducer. A biotinylated PM (b-PM) coated electrode that is layered with protein A-oriented antibodies against hemoglobin (Hb) readily captures non-glycated Hb (HbA0) and generates less photocurrent. The spectra of bacteriorhodopsin and Hb overlap so the photocurrent is reduced because of the partial absorption of the incident light by the captured Hb molecules. Two HbA0 and HbA1c aptasensors that are prepared by conjugating specific aptamers on b-PM coated electrodes single-step detect HbA0 and HbA1c in 15 min, without cross reactivity, with detection limits of ≤0.1 μg/mL and a dynamic range of 0.1–100 μg/mL. Both aptasensors exhibit high selectivity and long-term stability. For the clinical samples, HbA0 concentrations and HbA1c levels that are measured with aptasensors correlate well with total Hb concentrations and the HbA1c levels that are determined using standard methods (correlation gradient = 0.915 ± 0.004 and 0.981 ± 0.001, respectively). The use of these aptasensors for diabetes care is demonstrated. MDPI 2020-12-18 /pmc/articles/PMC7765918/ /pubmed/33353006 http://dx.doi.org/10.3390/s20247274 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lin, Ying-Chin
Lin, Ching-Yu
Chen, Hsiu-Mei
Kuo, Li-Pin
Hsieh, Cheng-En
Wang, Xiang-He
Cheng, Chih-Wen
Wu, Chih-Yin
Chen, Yi-Su
Direct and Label-Free Determination of Human Glycated Hemoglobin Levels Using Bacteriorhodopsin as the Biosensor Transducer
title Direct and Label-Free Determination of Human Glycated Hemoglobin Levels Using Bacteriorhodopsin as the Biosensor Transducer
title_full Direct and Label-Free Determination of Human Glycated Hemoglobin Levels Using Bacteriorhodopsin as the Biosensor Transducer
title_fullStr Direct and Label-Free Determination of Human Glycated Hemoglobin Levels Using Bacteriorhodopsin as the Biosensor Transducer
title_full_unstemmed Direct and Label-Free Determination of Human Glycated Hemoglobin Levels Using Bacteriorhodopsin as the Biosensor Transducer
title_short Direct and Label-Free Determination of Human Glycated Hemoglobin Levels Using Bacteriorhodopsin as the Biosensor Transducer
title_sort direct and label-free determination of human glycated hemoglobin levels using bacteriorhodopsin as the biosensor transducer
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7765918/
https://www.ncbi.nlm.nih.gov/pubmed/33353006
http://dx.doi.org/10.3390/s20247274
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