Beyond Sensitive and Selective Electrochemical Biosensors: Towards Continuous, Real-Time, Antibiofouling and Calibration-Free Devices
Abstract
:1. Introduction
2. Continuous, Real-Time Electrochemical Biosensors: Towards Antibiofouling, Reagentless, No-Wash, Single-Step, Reusable, and Calibration-Free Devices
2.1. Electrochemical Biosensors with Antibiofouling Properties
2.2. Reagentless, No-Wash, Single-Step, Near Real-Time, and Reusable Electrochemical Biosensors
2.3. Calibration-Free Biosensors
3. Opportunities, Impact, Challenges, and Future Insights
- Able to achieve high sensitivity and selectivity when defied punctually in multicomponent and protein-rich samples or continuously in flowing undiluted samples.
- Capable of responding to ups and downs in analyte concentration within seconds or minutes in a reversible way and without batch processing or addition of exogenous reagents.
- Insensitive to biofouling and stable after storage for more than one week in room-temperature blood serum.
- In ingestible formats coupled to transitory commercial polymer coatings with remarkable prolonged resistance to complex media of denaturing pH values such as gastrointestinal fluids.
- Integrated into microfluidic systems to monitor cell secretions.
- Deployed on screen-printed electrodes to provide rapid and accurate determinations when challenged to in finger-prick-scale sample volumes, suitable for application in POCT circumstances.
- Able to minimize the variability of the sensors fabrication and baseline drift and provide the required accuracy when operating continuously in vivo without the need for calibration, invoking physical barriers or using active drift-correction algorithms, thus surpassing main limitations of the commercial continuous glucose monitors.
Author Contributions
Funding
Conflicts of Interest
References
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Electrode | Sensor Fundamentals | Transduction Technique | Attribute (used Approach) | Additional Features | Molecular Target Tested | L.R./LOD | Sample | Ref. |
---|---|---|---|---|---|---|---|---|
16× Au electrode arrays prepared by photolithography | Sandwich hybridization assay at arrays modified with SHCP/HDT+MCH | Chrono-amperometry (TMB/H2O2) | Antibiofouling (thiolated ternary monolayer) | — | Synthetic target DNA (characteristic region of E. coli 16S rRNA) | LOD: 7 pM and 17 pM in spiked undiluted human serum and urine | Raw undiluted human serum and urine | [20] |
Au/SPEs | Sandwich hybridization assay at arrays modified with SHCP/HDT+MCH | Chrono-amperometry (TMB/H2O2) | Antibiofouling (thiolated ternary monolayer) | — | Synthetic target DNA and E. coli 16S rRNA | LOD: 25 pM and 100 pM in spiked undiluted human serum and urine and 16S rRNA E. coli corresponding to 3000 CFU mL−1 in raw cell lysate samples | Untreated raw serum, urine, and crude bacterial lysate solutions | [21] |
AuE | E-AB: Aptamer dually modified with a thiol and a redox reporter + PC-terminated SAM | SWV (MB) | Antibiofouling (PC-terminated SAM) | Continuous operation label-free | Kanamycin, doxorubicin | — | Flowing whole blood, both in vitro and in vivo (sensors placed in the jugular veins of live rats) | [12] |
GOx-PB-graphite SPEs | Electrode modified with Eudragit® L100 | CV ([Fe(CN)6]4−/3−) and Chrono-amperometry (PB/H2O2) | Antibiofouling (pH-sensitive transient polymer coating) | Continuous operation | Glucose | — | Raw undiluted blood and saliva | [22] |
Edible carbon paste GOx biosensors | Electrodes coated with Eudragit® E PO (pH < 5.0) or Eudragit® L100 (pH > 6.0) | Chrono-amperometry (PB/H2O2) | Antibiofouling (pH-sensitive transient polymer coating) | Continuous operation Biocatalytic activity preservation at media with denaturing pH values | Glucose | L.R.: 2−10 mM | GI fluids | [23] |
PEDOT-citrate film-modified GCE | Covalent immobilization using EDC/NHS chemistry of a peptide with anchoring, antifouling, and recognizing capabilities onto GCE/PEDOT-citrate | DPV ([Fe(CN)6]4−/3−) | Antibiofouling (multifunctional peptide) | — | APN, HepG2 cells | L.R.: 1 ng mL−1−15 μg mL−1 (APN) and 50–5 × 105 cells mL−1 (HepG2 cells) LOD: 0.4 ng mL−1 (APN) and 20 cells mL−1 (HepG2 cells) | Human urine | [24] |
Au disk | E-DNA: DNA probe dually modified with a thiol and a redox reporter + MCH SAM | SWV (MB) | Continuous and real-time operation (Folded-biosensor) | Reagentless and single-step | Melamine | LOD: 150 μM (∼19 ppm) in buffered solutions and 20 μM (∼2.5 ppm) in whole milk | Flowing undiluted whole milk | [6] |
Au | E-DNA: TDNs with two functional DNA/aptamer strands, one of them modified with MB | SWV (MB) | Continuous and real-time operation (Folded-biosensor) | Reagentless and single-step Antibiofouling Reusability | Target DNA, ATP | LOD: 300 fM (target DNA), 5 nM (ATP) | Flowing whole blood | [8] |
AuE | E-DNA: nucleic acid “scaffold” attached on one end to an electrode and presenting both a redox reporter and a specific epitope on the other | SWV (MB) | Reagentless and single-step (Folded-biosensor) | — | Three types of HIV-diagnostic antibodies | — | Human serum | [25] |
Microfabricated Au onto MECAS chip | E-AB: Aptamer dually modified with a thiol and a redox reporter + MCH SAM | ACV (MB) | Continuous and real-time operation (Folded-biosensor) | Reagentless and single-step Reusability | Cocaine | — | Flowing undiluted blood serum | [9] |
100 nm Au layer sputtered on glass slides | E-AB: Hairpin structure aptamer dually modified with a thiol and a redox reporter (MB or AQ) + MCH SAM | SWV (MB, AQ) | Continuous operation (Folded-biosensor) | Antibiofouling Reagentless and single-step | IFN-γ + TNF-α | LOD: 6.35 ng mL−1 (IFN-γ), 5.46 ng mL−1 (TNF-α) | Integrated into microfluidic devices to dynamically monitoring of cytokine release from immune cells (2.5 h) | [26] |
Au wire | E-AB: Aptamer dually modified with a thiol and a redox reporter + MCH SAM | SWV (MB) | Continuous and real-time and in vivo operation (Folded-biosensor) | Reagentless and single-step | Doxorubicin, Kanamycin, Gentamycin, and Tobramycin | — | Bloodstream awake, ambulatory rats | [10] |
Au disk, Au-plated SPCEs | E-PB: Peptide dually modified with a thiol and a redox reporter + MCH SAM | ACV, CV (MB) | Real-time operation (Folded-biosensor) | Reagentless and single-step | Pb2+ | LOD: 5 μM (ACV) | Diluted tap water, saliva, and urine samples | [27] |
Au disk | E-ION: T-rich ssDNA dually modified with thiol and redox reporter + Hg2+ + MCH SAM | ACV (MB) | Real-time operation (Folded-biosensor) | Reagentless and single-step Reusable | GSH (displaces Hg2+ by chelation) | LOD: 5 nM | 50% synthetic human saliva | [28] |
AuE | E-AB: Aptamer dually modified with a thiol and a redox reporter + MCH SAM | SWV (MB) | Calibration-free (“dual-frequency”) | Continuous and real-time operation Reagentless and single-step | Cocaine, doxorubicin | — | Continuous measurement in flowing, undiluted whole blood | [7] |
Au-SPE | E-AB: Aptamer dually modified with a thiol and a redox reporter + MCH | SWV (MB) | Calibration-free (“dual-frequency”) | Reagentless and single-step | Phenylalanine | L.R.: 90 nM−7 μM | Whole blood (diluted 1000-fold to match the sensor’s dynamic range) | [3] |
AuE | E-AB: Aptamer modified with a thiol and two different redox reporters + PC-terminated SAM | SWV (MB and AQ) | Calibration-free, (“dual reporter”) and in vivo operation | Continuous operation Antibiofouling Reagentless and single-step | Cocaine, ATP, kanamycin | — | Flowing whole blood, both in vitro and in vivo (sensors placed in the jugular veins of live rats) | [2] |
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Campuzano, S.; Pedrero, M.; Gamella, M.; Serafín, V.; Yáñez-Sedeño, P.; Pingarrón, J.M. Beyond Sensitive and Selective Electrochemical Biosensors: Towards Continuous, Real-Time, Antibiofouling and Calibration-Free Devices. Sensors 2020, 20, 3376. https://doi.org/10.3390/s20123376
Campuzano S, Pedrero M, Gamella M, Serafín V, Yáñez-Sedeño P, Pingarrón JM. Beyond Sensitive and Selective Electrochemical Biosensors: Towards Continuous, Real-Time, Antibiofouling and Calibration-Free Devices. Sensors. 2020; 20(12):3376. https://doi.org/10.3390/s20123376
Chicago/Turabian StyleCampuzano, Susana, María Pedrero, Maria Gamella, Verónica Serafín, Paloma Yáñez-Sedeño, and José Manuel Pingarrón. 2020. "Beyond Sensitive and Selective Electrochemical Biosensors: Towards Continuous, Real-Time, Antibiofouling and Calibration-Free Devices" Sensors 20, no. 12: 3376. https://doi.org/10.3390/s20123376