Hybrid Impedimetric Biosensors for Express Protein Markers Detection

Abstract

Impedimetric biosensors represent a powerful and promising tool for studying and monitoring biological processes associated with proteins and can contribute to the development of new approaches in the diagnosis and treatment of diseases. The basic principles, analytical methods, and applications of hybrid impedimetric biosensors for express protein detection in biological fluids are described. The advantages of this type of biosensors, such as simplicity and speed of operation, sensitivity and selectivity of analysis, cost-effectiveness, and an ability to be integrated into hybrid microfluidic systems, are demonstrated. Current challenges and development prospects in this area are analyzed. They include (a) the selection of materials for electrodes and formation of nanostructures on their surface; (b) the development of efficient methods for biorecognition elements' deposition on the electrodes' surface, providing the specificity and sensitivity of biosensing; (c) the reducing of nonspecific binding and interference, which could affect specificity; (d) adapting biosensors to real samples and conditions of operation; (e) expanding the range of detected proteins; and, finally, (f) the development of biosensor integration into large microanalytical system technologies. This review could be useful for researchers working in the field of impedimetric biosensors for protein detection, as well as for those interested in the application of this type of biosensor in biomedical diagnostics.

Keywords: antibodies; aptamers; electrochemical impedance spectroscopy; express detection; impedimetric biosensors; label-free detection; microfluidics; nanomaterials; peptides; proteins.

Figure 1

A general scheme for the development of a hybrid impedimetric biosensor for protein detection. 1—selecting the design and material of electrodes; 2—electrode modification and functionalization; 3—selection and immobilization of biorecognition element; 4—implementation of the interface with measurement circuit and sample analysis system.

Figure 2

Typical Nyquist diagram (a) with corresponding equivalent electrical circuit (b).

Figure 3

Schematic example of a Bode plot for impedance module, |Z|, and phase shift versus frequency, f.

Figure 4

Schematic presentation of immunosensor preparation process for SPB detection. Reprinted from [68] with permission of Elsevier provided by Copyright Clearance Center.

Figure 5

Scheme of an immunosensor for PAK 2 based on an ITO electrode. Reprinted from [76] with permission of Elsevier provided by Copyright Clearance Center.

Figure 6

ST2 immunosensor immobilization scheme. Reprinted from [88] with permission of Elsevier provided by Copyright Clearance Center.

Figure 7

SEM micro-images of the bare B:CNW surface. Reprinted from [99] with permission of Elsevier provided by Copyright Clearance Center.

Figure 8

SEM image with low and high magnification (a,b), TEM (c), and high-resolution TEM (d) images of HsGDY@NDs. Reprinted from [100] with permission of Elsevier provided by Copyright Clearance Center.

Figure 9

Schematic presentation of manufacturing steps of a biosensor with a Au electrode: (A)—Au electrode without functionalization; (B)—adsorption of cysteamine on the gold surface; (C) covalent binding of carboxybetaine monomer (CBMA) in the presence of EDC/NHS for activation; (D)—photopolymerization of the CBMA monomer in solution led to the formation of a zwitterionic polymer pCBMA with a CBMA-receptive interface for covalent binding of the IL-8 polyclonal antibody (Ab) in the presence of EDC/NHS (E) to capture the target analyte (F). Reprinted from [102] with permission of Elsevier provided by Copyright Clearance Center.

Figure 10

FE-SEM images of electropolymerized coatings of FTO/PPy (A–C), FTO/PPy-MO NIP (D–F), and FTO/PPy-MO DMIP electrodes (G–I). The DMIP electrode was prepared by imprinting 100 ng/mL of each biomarker AFP and CEA. Reprinted from [113] with permission of Elsevier provided by Copyright Clearance Center.

Figure 11

SEM images of electrode modifications AuE (A), AuE/4-ATP/fullerene (B), and AuE/4-ATP/fullerene/PAMAM (C). Reprinted from [131] with permission of Elsevier provided by Copyright Clearance Center.

Figure 12

The original nickel foam and the resulting FeCo-MOF/NF: (a) SEM images of bare porous NF, (b–e) FeCo-MOF grown on the NF skeleton (FeCo-MOF/NF) after in situ hydrothermal reactions using precursors, ligands, and Ni foam skeleton at different magnifications. The marks in (f) are the average thickness of FeCo-MOF nanosheets grown on porous NF substrates. Reprinted from [134] with permission of Elsevier provided by Copyright Clearance Center.

Figure 13

SEM images of top view (A,B) and cross-view (C,D) of TNT (A,C) and TNT annealed in argon (B,D) at 550 °C for 2 h. Reprinted from [141], license CC BY-NC-ND 4.0.

Figure 14

Non-faradaic impedimetric biosensor monitoring of caspase-9 in mammalian cell culture: (a) SEM image of a film of ZnO/CuO nanoparticles; (b) illustration of a cross-section of the biosensor surface and biorecognition interactions between anti-cas9 and caspase-9. Reprinted from [51], license CC BY-NC-ND 4.0.

Figure 15

Schematic representation of the stages of immunosensor formation with QD for detection of EP-ZIKV. Reprinted from [153] with permission of Elsevier provided by Copyright Clearance Center.

Figure 16

SEM electron microscopy images of MoS₂ nanoflowers: (a) flower like morphology; (b) single bud with a more magnified petals. Reprinted from [154] with permission of Elsevier provided by Copyright Clearance Center.

Figure 17

Modification stages of Au electrode using aryldiazonium (CB) carboxybetaine derivative and lectin (SNA-I). Reprinted from [157] with permission of Elsevier provided by Copyright Clearance Center.

Figure 18

COVID-19 impedimetric biosensor based on polypyrrole nanotubes, nickel hydroxide, and VHH antibody fragment. Reprinted from [210] with permission of Elsevier provided by Copyright Clearance Center.

Figure 19

Preparation of L-cysteine-SnTeSe QD aptasensor. Reprinted from [152] with permission of Elsevier provided by Copyright Clearance Center.

Figure 20

Analytical performance for the detection of NGAL: a commercially available ELISA NGAL detection kit (a) compared to sensor system from [230] (b). Statistical significance between groups was determined by ANOVA test, where p value is * p < 0.02 or ** p < 0.256 in (a), * p < 0.017 or ** p < 0.246 in (b), respectively. Mean values of each group are indicated with bold lines. Reprinted from [230] with permission of Elsevier provided by Copyright Clearance Center.

Figure 21

(a) Fabrication process of the (NGAL-BP1)-modified PDMS mold with hexagonally patterned pores for peptide surface imprinting via incubation in a peptide solution with a specific concentration. (b) Atomic force microscopy (AFM) image of the polystyrene (PS) colloidal monolayer and (c,d) scanning electron microscopy (SEM) images of oxidized PDMS molds with patterned concave pores after NGAL-BP1 adsorption during 2.5 h incubation in 9.25 and 18.51 μg/mL peptide solutions. All scale bars are 1 µm. Reprinted from [124] with permission of Elsevier provided by Copyright Clearance Center.

Figure 22

Molecularly imprinted polymer-based electrochemical impedimetric sensor for detection of trace cytokine IL-1β. Reprinted from [127] with permission of Elsevier provided by Copyright Clearance Center.

Figure 23

Schematic drawing of conceptual basis of IL-6-recognizing biosensor. Gold electrodes are modified with a nanobody or an aptamer specifically recognizing IL-6. There is a change in the non-faradaic impedance spectra when these surfaces are challenged in serum samples containing IL-6, which can be observed in both impedance (a) and capacitance planes (b). Reprinted from [247], license CC BY-NC-ND 4.0.

Figure 24

The schematic illustration of the immunosensor preparation based on gold nanostructure (AuNS) electrodeposition followed by 3-mercaptopropionic acid (MPA) modification, antibody immobilization, and BSA blocking (A). The design and assembly of a microfluidic device integrating top–bottom opposite electrodes (B). The device was connected to a syringe pump for controlling the flow rate and an electrochemical apparatus for EIS measurement (C). Reprinted from [267] with permission of Elsevier provided by Copyright Clearance Center.

Figure 25

A 3D view of the biosensor CHD for diagnosis. Reprinted from [271], license CC BY-NC-ND 4.0.