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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Bibliographic Details
Published in:Sensors (Basel, Switzerland) Switzerland), 2020-12, Vol.20 (24), p.7274
Main Authors: 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
Format: Article
Language:English
Subjects:
Animals
Antibodies
Bacteriorhodopsin
Bacteriorhodopsins
Biosensing Techniques
Biosensors
Capillary electrophoresis
Chromatography
Coated electrodes
Correlation analysis
Diabetes
Electrodes
Glucose
glycated hemoglobin
Glycated Hemoglobin A - analysis
Graphene
Hemoglobin
Humans
Incident light
photoelectric biosensor
Photoelectric effect
Photoelectric emission
Photoelectricity
Protein A
Proteins
purple membrane
Rabbits
Reynolds number
Selectivity
Sucrose
Transducers
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Summary: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.
ISSN:1424-8220
1424-8220
DOI:10.3390/s20247274