Construction and optimization of a versatile bioFET platform for detecting nucleic acids and protein using the biotin–neutravidin affinity system

A biosensor based on a field-effect transistor (bioFET) is a promising system for point-of-care testing due to its portability and high sensitivity. A bioFET uses a linker platform to connect capture molecules to the FET, which detects electrical changes when targets bind to the capture molecules. T...

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Main Authors: Jiunn-Tyng Yeh, Lian-Chin Wang, Chi-Pei Weng, Chang-Fu Kuo
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Sensing and Bio-Sensing Research
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Online Access:http://www.sciencedirect.com/science/article/pii/S2214180425000443
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author Jiunn-Tyng Yeh
Lian-Chin Wang
Chi-Pei Weng
Chang-Fu Kuo
author_facet Jiunn-Tyng Yeh
Lian-Chin Wang
Chi-Pei Weng
Chang-Fu Kuo
author_sort Jiunn-Tyng Yeh
collection DOAJ
description A biosensor based on a field-effect transistor (bioFET) is a promising system for point-of-care testing due to its portability and high sensitivity. A bioFET uses a linker platform to connect capture molecules to the FET, which detects electrical changes when targets bind to the capture molecules. This study presents the development and optimization of a versatile, highly sensitive bioFET platform utilizing a biotin–neutravidin affinity system capable of detecting both DNA and proteins. The bioFET was constructed using an extended-gate field-effect transistor (EGFET) with an aluminum gate functionalized with (3-aminopropyl)triethoxysilane (APTES)–biotin–neutravidin. We optimized the concentrations and molar ratio of biotin and neutravidin using fluorescence imaging and biosensing benchmarks. We found that 0.1 μg/mL biotin paired with 30 μg/mL neutravidin, approximately 1:1 in molarity, achieved optimal sensitivity. The optimized bioFET system demonstrated good sensitivity, with lower detection limits of 3.5 copies for Escherichia coli genomic DNA and 0.3 fg/mL for p-Tau217, an Alzheimer's disease biomarker. In summary, this study establishes the ideal conditions for constructing a bioFET system using an APTES–biotin–neutravidin linker capable of detecting DNA and proteins, with potential for convenient and sensitive point-of-care diagnostics.
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publishDate 2025-06-01
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series Sensing and Bio-Sensing Research
spelling doaj-art-b80153f735d348bdb8e5b02bcd5e1d6d2025-08-20T03:20:03ZengElsevierSensing and Bio-Sensing Research2214-18042025-06-014810077810.1016/j.sbsr.2025.100778Construction and optimization of a versatile bioFET platform for detecting nucleic acids and protein using the biotin–neutravidin affinity systemJiunn-Tyng Yeh0Lian-Chin Wang1Chi-Pei Weng2Chang-Fu Kuo3Novascope Diagnostics INC, Taipei, Taiwan; Center for Artificial Intelligence in Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, TaiwanNovascope Diagnostics INC, Taipei, TaiwanNovascope Diagnostics INC, Taipei, TaiwanNovascope Diagnostics INC, Taipei, Taiwan; Center for Artificial Intelligence in Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan; Division of Rheumatology, Allergy and Immunology, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan; Department of Internal Medicine, College of Medicine, Chang Gung University, Taoyuan, Taiwan; Division of Rheumatology, Orthopaedics and Dermatology, School of Medicine, University of Nottingham, Nottingham, UK; Corresponding author at: Research Building 3rd Floor AI Core Lab, No. 15 Wenhua 1st Rd., Guishan Dist., Taoyuan 333, Taiwan.A biosensor based on a field-effect transistor (bioFET) is a promising system for point-of-care testing due to its portability and high sensitivity. A bioFET uses a linker platform to connect capture molecules to the FET, which detects electrical changes when targets bind to the capture molecules. This study presents the development and optimization of a versatile, highly sensitive bioFET platform utilizing a biotin–neutravidin affinity system capable of detecting both DNA and proteins. The bioFET was constructed using an extended-gate field-effect transistor (EGFET) with an aluminum gate functionalized with (3-aminopropyl)triethoxysilane (APTES)–biotin–neutravidin. We optimized the concentrations and molar ratio of biotin and neutravidin using fluorescence imaging and biosensing benchmarks. We found that 0.1 μg/mL biotin paired with 30 μg/mL neutravidin, approximately 1:1 in molarity, achieved optimal sensitivity. The optimized bioFET system demonstrated good sensitivity, with lower detection limits of 3.5 copies for Escherichia coli genomic DNA and 0.3 fg/mL for p-Tau217, an Alzheimer's disease biomarker. In summary, this study establishes the ideal conditions for constructing a bioFET system using an APTES–biotin–neutravidin linker capable of detecting DNA and proteins, with potential for convenient and sensitive point-of-care diagnostics.http://www.sciencedirect.com/science/article/pii/S2214180425000443EGFETbioFETBiotinNeutravidinBiosensor
spellingShingle Jiunn-Tyng Yeh
Lian-Chin Wang
Chi-Pei Weng
Chang-Fu Kuo
Construction and optimization of a versatile bioFET platform for detecting nucleic acids and protein using the biotin–neutravidin affinity system
Sensing and Bio-Sensing Research
EGFET
bioFET
Biotin
Neutravidin
Biosensor
title Construction and optimization of a versatile bioFET platform for detecting nucleic acids and protein using the biotin–neutravidin affinity system
title_full Construction and optimization of a versatile bioFET platform for detecting nucleic acids and protein using the biotin–neutravidin affinity system
title_fullStr Construction and optimization of a versatile bioFET platform for detecting nucleic acids and protein using the biotin–neutravidin affinity system
title_full_unstemmed Construction and optimization of a versatile bioFET platform for detecting nucleic acids and protein using the biotin–neutravidin affinity system
title_short Construction and optimization of a versatile bioFET platform for detecting nucleic acids and protein using the biotin–neutravidin affinity system
title_sort construction and optimization of a versatile biofet platform for detecting nucleic acids and protein using the biotin neutravidin affinity system
topic EGFET
bioFET
Biotin
Neutravidin
Biosensor
url http://www.sciencedirect.com/science/article/pii/S2214180425000443
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