Wearable sweat biosensors offer compelling opportunities for improved personal health monitoring and non-invasive measurements of key biomarkers. Inexpensive device fabrication methods are necessary for scalable manufacturing of portable, disposable, and flexible sweat sensors. Furthermore, real-time sweat assessment must be analyzed to validate measurement reliability at various sweating rates. Here, we demonstrate a “smart bandage” microfluidic platform for cortisol detection and continuous glucose monitoring integrated with a synthetic skin. The low-cost, laser-cut microfluidic device is composed of an adhesive-based microchannel and solution-processed electrochemical sensors fabricated from inkjet-printed graphene and silver solutions. An antibody-derived cortisol sensor achieved a limit of detection of 10 pM and included a low-voltage electrowetting valve, validating the microfluidic sensor design under typical physiological conditions. To understand effects of perspiration rate on sensor performance, a synthetic skin was developed using soft lithography to mimic human sweat pores and sweating rates. The enzymatic glucose sensor exhibited a range of 0.2 to 1.0 mM, a limit of detection of 10 μM, and reproducible response curves at flow rates of 2.0 μL min −1 and higher when integrated with the synthetic skin, validating its relevance for human health monitoring. These results demonstrate the potential of using printed microfluidic sweat sensors as a low-cost, real-time, multi-diagnostic device for human health monitoring.
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An integrated microfluidic platform for selective and real-time detection of thrombin biomarkers using a graphene FET
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.
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- Award ID(s):
- 1847152
- NSF-PAR ID:
- 10215343
- Date Published:
- Journal Name:
- The Analyst
- Volume:
- 145
- Issue:
- 13
- ISSN:
- 0003-2654
- Page Range / eLocation ID:
- 4494 to 4503
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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null (Ed.)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
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null (Ed.)An ultrasensitive and versatile assay for biomarkers has been developed using graphene/gold nanoparticles (AuNPs) composites and single-particle inductively-coupled plasma/mass spectrometry (spICP-MS). Thrombin was chosen as a model biomarker for this study. AuNPs modified with thrombin aptamers were first non-selectively adsorbed onto the surface of graphene oxide (GO) to form GO/AuNPs composites. In the presence of thrombin, the AuNPs desorbed from the GO/AuNPs composites due to a conformation change of the thrombin aptamer after binding with thrombin. The desorbed AuNPs were proportional to the concentration of thrombin and could be quantified by spICP-MS. By counting the individual AuNPs in the spICP-MS measurement, the concentration of thrombin could be determined. This assay achieved an ultralow detection limit of 4.5 fM with a broad linear range from 10 fM to 100 pM. The method also showed excellent selectivity and reproducibility when a complex protein matrix was evaluated. Furthermore, the diversity and ready availability of ssDNA ligands make this method a versatile new technique for ultrasensitive detection of a wide variety of biomarkers in clinical diagnostics.more » « less
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Biosensors based on graphene field effect transistors (GFETs) decorated with antibody‐functionalized platinum nanoparticles (PtNPs) are developed for the quantitative detection of breast cancer biomarker HER3. High‐quality chemical vapor deposited graphene is prepared and transferred over gold electrodes microfabricated on an SiO2/Si wafer to yield an array of 52 GFET devices. The GFETs are modified with PtNPs to obtain a hybrid nanostructure suitable for attachment of HER3‐specific, genetically engineered thiol‐containing single‐chain variable fragment antibodies (scFv) to realize a biosensor for HER3. Physical and electrical characterization of Bio‐GFET devices is carried out by electron microscopy, atomic force microscopy, Raman spectroscopy, and current–gate voltage measurements. A concentration‐dependent response of the biosensor to HER3 antigen is found in the range 300 fg mL−1to 300 ng mL−1and is in quantitative agreement with a model based on the Hill–Langmuir equation of equilibrium thermodynamics. Based on the dose–response data, the dissociation constant is estimated to be 800 pg mL−1, indicating that the high affinity of the scFv antibody is maintained after immobilization. The limit of detection is 300 fg mL−1, showing the potential for PtNP/G‐FETs to be used in label‐free biological sensors.
- Free Publicly Accessible Full Text
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.1039/D0AN00251H
