skip to main content
US FlagAn official website of the United States government
dot gov icon
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
https lock icon
Secure .gov websites use HTTPS
A lock ( lock ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.


Title: Detection of an IL-6 Biomarker Using a GFET Platform Developed with a Facile Organic Solvent-Free Aptamer Immobilization Approach
Aptamer-immobilized graphene field-effect transistors (GFETs) have become a well-known detection platform in the field of biosensing with various biomarkers such as proteins, bacteria, virus, as well as chemicals. A conventional aptamer immobilization technique on graphene involves a two-step crosslinking process. In the first step, a pyrene derivative is anchored onto the surface of graphene and, in the second step, an amine-terminated aptamer is crosslinked to the pyrene backbone with EDC/NHS (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride/N-hydroxysuccinimide) chemistry. However, this process often requires the use of organic solvents such as dimethyl formamide (DMF) or dimethyl sulfoxide (DMSO) which are typically polar aprotic solvents and hence dissolves both polar and nonpolar compounds. The use of such solvents can be especially problematic in the fabrication of lab-on-a-chip or point-of-care diagnostic platforms as they can attack vulnerable materials such as polymers, passivation layers and microfluidic tubing leading to device damage and fluid leakage. To remedy such challenges, in this work, we demonstrate the use of pyrene-tagged DNA aptamers (PTDA) for performing a one-step aptamer immobilization technique to implement a GFET-based biosensor for the detection of Interleukin-6 (IL-6) protein biomarker. In this approach, the aptamer terminal is pre-tagged with a pyrene group which becomes soluble in aqueous solution. This obviates the need for using organic solvents, thereby enhancing the device integrity. In addition, an external electric field is applied during the functionalization step to increase the efficiency of aptamer immobilization and hence improved coverage and density. The results from this work could potentially open up new avenues for the use of GFET-based BioMEMS platforms by broadening the choice of materials used for device fabrication and integration.  more » « less
Award ID(s):
1847152
PAR ID:
10215337
Author(s) / Creator(s):
;
Date Published:
Journal Name:
Sensors
Volume:
21
Issue:
4
ISSN:
1424-8220
Page Range / eLocation ID:
1335
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. null (Ed.)
    Lab-on-a-chip technology offers an ideal platform for low-cost, reliable, and easy-to-use diagnostics of key biomarkers needed for early screening of diseases and other health concerns. In this work, a graphene field-effect transistor (GFET) functionalized with target-binding aptamers is used as a biosensor for the detection of thrombin protein biomarker. Furthermore, this GFET is integrated with a microfluidic device for enhanced sensing performances in terms of detection limit, sensitivity, and continuous monitoring. Under this platform, a picomolar limit of detection was achieved for measuring thrombin; in our experiment measured as low as 2.6 pM. FTIR, Raman and UV-Vis spectroscopy measurements were performed to confirm the device functionalization steps. Based on the concentration-dependent calibration curve, a dissociation constant of K D = 375.8 pM was obtained. Continuous real-time measurements were also conducted under a constant gate voltage ( V GS ) to observe the transient response of the sensor when analyte was introduced to the device. The target selectivity of the sensor platform was evaluated and confirmed by challenging the GFET biosensor with various concentrations of lysozyme protein. The results suggest that this device technology has the potential to be used as a general diagnostic platform for measuring clinically relevant biomarkers for point-of-care applications. 
    more » « less
  2. Dementia is a brain disease which results in irreversible and progressive loss of cognition and motor activity. Despite global efforts, there is no simple and reliable diagnosis or treatment option. Current diagnosis involves indirect testing of commonly inaccessible biofluids and low-resolution brain imaging. We have developed a portable, wireless readout-based Graphene field-effect transistor (GFET) biosensor platform that can detect viruses, proteins, and small molecules with single-molecule sensitivity and specificity. We report the detection of three important amyloids, namely, Amyloid beta (Aβ), Tau (τ), and α-Synuclein (αS) using DNA aptamer nanoprobes. These amyloids were isolated, purified, and characterized from the autopsied brain tissues of Alzheimer’s Disease (AD) and Parkinson’s Disease (PD) patients. The limit of detection (LoD) of the sensor is 10 fM, 1–10 pM, 10–100 fM for Aβ, τ, and αS, respectively. Synthetic as well as autopsied brain-derived amyloids showed a statistically significant sensor response with respect to derived thresholds, confirming the ability to define diseased vs. nondiseased states. The detection of each amyloid was specific to their aptamers; Aβ, τ, and αS peptides when tested, respectively, with aptamers nonspecific to them showed statistically insignificant cross-reactivity. Thus, the aptamer-based GFET biosensor has high sensitivity and precision across a range of epidemiologically significant AD and PD variants. This portable diagnostic system would allow at-home and POC testing for neurodegenerative diseases globally. 
    more » « less
  3. We present a novel fabrication technique for large-scale, on-wafer graphene devices. With the proposed technique, large-area graphene apertures can be fabricated, enabling the proliferation of graphene-based reconfigurable devices, including metasurfaces. Such topologies require large-area high yield fabrication processes. To avoid graphene delamination during the chemical processes of the fabrication, we use a titanium sacrificial layer to protect the graphene monolayer. To evaluate the fabrication method, we present broadband in-plane graphene measurements in the 220-330 GHz band for the first time and compare the measured resistance sheet with previous works. 
    more » « less
  4. In the present study, hybrid nanoflowers (HNFs) based on copper (II) or manganese (II) ions were prepared by a simple method and used as nanosupports for the development of effective nanobiocatalysts through the immobilization of lipase B from Pseudozyma antarctica. The hybrid nanobiocatalysts were characterized by various techniques including scanning electron microscopy (SEM), energy dispersion spectroscopy (EDS), X-ray diffraction (XRD), Raman spectroscopy, and Fourier transform infrared spectroscopy (FTIR). The effect of the addition of carbon-based nanomaterials, namely graphene oxide and carbon nanotubes, as well as magnetic nanoparticles such as maghemite, on the structure, catalytic activity, and operational stability of the hybrid nanobiocatalysts was also investigated. In all cases, the addition of nanomaterials during the preparation of HNFs increased the catalytic activity and the operational stability of the immobilized biocatalyst. Lipase-based magnetic nanoflowers were effectively applied for the synthesis of tyrosol esters in non-aqueous media, such as organic solvents, ionic liquids, and environmental friendly deep eutectic solvents. In such media, the immobilized lipase preserved almost 100% of its initial activity after eight successive catalytic cycles, indicating that these hybrid magnetic nanoflowers can be applied for the development of efficient nanobiocatalytic systems. 
    more » « less
  5. null (Ed.)
    Graphene-based 3D macroscopic aerogels with their hierarchical porous structures and mechanical strength have been widely explored for removing contaminants from water. However, their large-scale manufacturing and application in various water treatment processes are limited by their scalability. In this study, we report a proof-of-concept direct ink writing (DIW) 3D printing technique and subsequent freeze-drying to prepare graphene-biopolymer aerogels for water treatment. To provide appropriate rheology for DIW printability, two bio-inspired polymers, polydopamine (PDA) and bovine serum albumin (BSA), were added to the graphene-based ink. The biopolymers also contributed to the contaminant removal capacity of the resultant graphene-polydopamine-bovine serum albumin (G-PDA-BSA) aerogel. The physicochemical properties of the aerogel were thoroughly characterized from the nano- to macroscale. The 3D printed aerogel exhibited excellent water contaminant removal performance for heavy metals (Cr( vi ), Pb( ii )), organic dyes (cationic methylene blue and anionic Evans blue), and organic solvents ( n -hexane, n -heptane, and toluene) in batch adsorption studies. The electrostatic interaction dominated the removal of heavy metals and dyes while the hydrophobic interaction dominated the removal of organic solvents from water. Moreover, the aerogel showed superb regeneration and reuse potential. The aerogel removed 100% organic solvents over 10 cycles of regeneration and reuse; additionally, the removal efficiencies for methylene blue decreased by 2–20% after the third cycle. The fit-for-design 3D printed aerogel was also effectively used as a bottle-cap flow-through filter for dye removal. The potential and vision of the 3D printing approach for graphene-based water treatment presented here can be extended to other functional nanomaterials, can enable shape-specific applications of fit-for-purpose adsorbents/reactors and point-of-use filters, and can materialize the large-scale manufacturing of nano-enabled water treatment devices and technologies. 
    more » « less