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Avidin-Biotin Technology in Gold Nanoparticle-Decorated Graphene Field Effect Transistors for Detection of Biotinylated Macromolecules with Ultrahigh Sensitivity and Specificity.

ABSTRACT: The strong and specific noncovalent interaction between avidin and biotin is widely exploited in different types of enzyme-linked immunosorbent assay kits, labeled immunosensors, and polymer-based sensing devices for the detection of different biomarkers specific to different diseases such as cancer and influenza. Here, we employed the avidin-biotin technology in a novel gold nanoparticle-decorated graphene field-effect transistor (AuNP-GFET) and demonstrated the specific detection of the biotinylated macromolecules such as biotinylated proteins and nucleotides in the sub-picomolar (pM) range. The AuNP-GFET was constructed by fabricating six pairs of interdigital electrodes on graphene transferred on a SiO₂/Si substrate. The sensing performance of AuNP-GFET was characterized by the real-time two-terminal electrical current measurement upon injection of the analyte solution into a silicone pool preattached onto the electrodes. Avidin, a tetrameric biotin-binding protein with strong affinity and specificity, immobilized on AuNP-decorated single-layer graphene, was used as the sensing platform and transduced the electrical signal upon binding to the analyte macromolecules. The sensing capability of the AuNP-GFET was tested with the biotinylated protein A. Sensitivity of the present biosensor was estimated to be ?0.4 pM. The specificity and applicability of the biosensor were confirmed using both synthetic and real samples. Because the biotin label can retain its binding capability to avidin with strong affinity and specificity even after conjugating with varieties of proteins and nucleotides, the present AuNP-GFET biosensor is expected to promote the research in developing different biosensors.

SUBMITTER: Wang S

PROVIDER: S-EPMC7689884 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature

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Avidin-Biotin Technology in Gold Nanoparticle-Decorated Graphene Field Effect Transistors for Detection of Biotinylated Macromolecules with Ultrahigh Sensitivity and Specificity.

Wang Shiyu S Hossain Md Zakir MZ Han Tao T Shinozuka Kazuo K Suzuki Takaaki T Kuwana Anna A Kobayashi Haruo H

ACS omega 20201112 46

The strong and specific noncovalent interaction between avidin and biotin is widely exploited in different types of enzyme-linked immunosorbent assay kits, labeled immunosensors, and polymer-based sensing devices for the detection of different biomarkers specific to different diseases such as cancer and influenza. Here, we employed the avidin-biotin technology in a novel gold nanoparticle-decorated graphene field-effect transistor (AuNP-GFET) and demonstrated the specific detection of the biotin ...[more]

PMID: 33251439

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Project description:Affinity interactions between the small molecule biotin and the protein avidin have been used extensively to functionalize biomaterials. More recently, researchers have leveraged the changes in biotin-avidin affinity that occur upon biotin conjugation to larger molecules to control the release of biotinylated drugs and proteins. However, the effects of biotin-avidin interactions on hydrogel properties have not been thoroughly investigated. The objective of this study was to evaluate the effect of increasing biotin and avidin concentrations on hydrogel swelling properties, as an indicator of crosslinking. Gelatin, selected as a model hydrogel material, was biotinylated at increasing fold molar excesses of biotin with a PEG linker using N-hydroxysuccinimide chemistry. Afterwards, biotinylated gelatin was formed into hydrogels and stabilized with glutaraldehyde. Swelling properties of the biotinylated hydrogels were investigated by conducting swelling studies in different avidin solutions. Increasing the degree of biotinylation caused significant decreases in swelling ratios of the hydrogels in a dose-dependent manner, suggesting increases in crosslinking of the hydrogels. However, increasing avidin concentrations in excess of biotin content did not significantly affect swelling ratios. Moving hydrogels to phosphate-buffered saline following avidin incorporation resulted in increased swelling ratios for hydrogels prepared with a lower concentration of biotin. However, hydrogels prepared with the highest concentration of biotin did not experience increased swelling ratios, implying that the stability of biotin-avidin-mediated crosslinking depends on the number of biotin molecules available for binding. Collectively, these results demonstrate that biotin-avidin interactions control hydrogel swelling properties, and that the magnitude and stability of the effects depend on the biotin concentration. These results have important implications for affinity-based controlled release of biotinylated drugs or proteins from biotin-avidin-crosslinked hydrogels.

Dataset Information

Avidin-Biotin Technology in Gold Nanoparticle-Decorated Graphene Field Effect Transistors for Detection of Biotinylated Macromolecules with Ultrahigh Sensitivity and Specificity.

Publications

Avidin-Biotin Technology in Gold Nanoparticle-Decorated Graphene Field Effect Transistors for Detection of Biotinylated Macromolecules with Ultrahigh Sensitivity and Specificity.

Similar Datasets

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