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SARS-CoV-2 biosensor fabrication steps.

Background: Biomarkers are very important disease-related biomolecules which should be analyzed sensitive and selective in related physiological fluids or tissues. Tumor necrosis factor-α is a type of cytokine which plays vitlly important roles in different methabolic pathways such as cell death, survival, differentiation, proliferation and migration, and infectious and inflammatory diseases including rheumatoid arthritis, diabetes. Objective: In this study, it was aimed to develop a reliable tool based on star-shaped poly(glycidyl methacrylate) polymer coated disposable indium tin oxide electrode for determination of Tumor necrosis factor-α, an important disease biomarker. Methods: Star shaped polymer was used as an interface material for anti- Tumor necrosis factor α antibodies immobilization. The antibodies were immobilized covalently onto polymer coated indium tin oxide electrode. Electrochemical impedance spectroscopy and cyclic voltammetry techniques were used for all electrochemical measurements. Results: The suggested immunosensor exhibited a linear range between 0.02 and 4 pg/mL Tumor necrosis factor-α, and the detection limit was found as 6 fg/mL. Scanning electron microscopy and atomic force microscopy were used for electrode surface characterization. In addition, the suggested immunosensor was used for Tumor necrosis factor-α sensing in human serum samples. The results displayed recoveries between 97.07 and 100.19%. Moreover, this immunosensor had a simple fabrication procedure and a long storage-stability. Conclusion: A new biosensor based on a Star shaped polymer for the ultra sensitive determination of a biomarker Tumor necrosis factor-α was developed. The biosensor presented excellent repeatability and reproducubility, and also wide calibration range for Tumor necrosis factor- α.

A label-free impedimetric immunosensing system was constructed for ultrasensitive determination of the calreticulin (CALR) biological marker in human serum samples utilizing an electrochemical impedance spectroscopy analysis technique for the first time. The new biosensor fabrication procedure consisted of electrodeposition of single-walled carbon nanotubes (SWCNTs) incorporating polymerization of an oxiran-2-yl methyl 3-(1H-pyrrol-1-yl) propanoate monomer (

In this study, we constructed a disposable indium tin oxide polyethylene terephthalate film (ITO-PET)-based electrochemical aptasensor for analyzing prostate-specific antigen (PSA), one of the most important biomarkers of prostate cancer. Because of their clinical importance, building PSA biosensing systems with high sensitivity and stability is essential. However, it still presents significant difficulties, such as low detection limits. We designed a platform to covalently bind the amino-terminated aptamer by modifying the ITO-PET surface with carboxyethylsilanetriol (CTES) to obtain a self-assembled monolayer (SAM). We also evaluated the potential for use in real human serum samples by investigating the optimal operating conditions and analytical performance characteristics of the developed biosensor. The design we present here exhibits excellent precision, with a limit of detection (LOD) as low as 8.74 fg/mL PSA. The broad linear detection range of the biosensor under optimal conditions was determined as 1.0-1500 fg/mL. The dissociation constant (K

A new impedimetric biosensing system based on kallikrein-related peptidase 4 (KLK 4) antigen-specific antibodies and a zinc(II) phthalocyanine tetracarboxylic acid (Zn-PcTCa) matrix material was developed for the first time in this study. First, a Zn-PcTCa-coated indium tin oxide surface was used as an interface matrix material for the immobilization of anti-KLK 4 antibodies, and they bound to the platform via amide bonds. In the presence of KLK 4 antigens, the anti-KLK 4 antibodies specifically captured these antigens and caused changes in the electrochemical properties of the system. Randles equivalent circuit was utilized to evaluate the impedimetric signal, which was measured with the help of an electrochemical impedance spectroscopy method. After the specific interaction, the electron transfer resistance (Rct) was remarkably increased and displayed a linear relationship with the level of the KLK 4 antigen in the range of 0.02-15 pg/mL, with a a detection limit of 6.8 fg/mL. The designed biosensor was able to detect a KLK 4 antigen with good sensitivity, excellent specificity, and high stability. In addition, because of having a low-cost and robust procedure for fabrication, it could be repeatedly used in several areas including clinical diagnosis.

In present study, we developed a highly sensitive, electrochemical immunosensor based on fullerene C60-modified disposable graphite paper (GP) electrode for determination of Suppression of Tumorigenicity 2 (ST2) in human serum. The synthesis of the ST2 immunosensor was monitored with electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) techniques and single frequency impedance (SFI) technique which is utilized for the specific interaction between anti-ST2 and ST2 antigen. Moreover, the morphological alteration of each GP surface was examined by scanning electron microscopy (SEM), SEM-energy dispersive X-ray spectroscopy (EDX) and atomic force microscopy (AFM). All parameters such as fullerene C60 concentration, antibody concentration and antibody incubation time were optimized. Analytical characteristics such as linear determination range, repeatability, reproducibility, regeneration and surface coverage were determined for the immunosensor. The ST2 electrochemical immunosensor had excellent repeatability, reproducibility and a wide detection range (from 0.1 fg mL−1 to 100 fg mL−1). The proposed immunosensor also had low limit of detection (LOD) and limit of quantification (LOQ) values of 0.124 fg mL−1 and 0.414 fg mL−1, respectively. The proposed immunosensor was applied to real samples to test applicability in clinical practice.

In this study we synthesized Pd(0) nanoparticles with nearly 20 nm size using p(3-Sulfopropyl acrylate) (p(SPA)) hydrogel as support material. Pd@p(SPA) containing Pd (0) nanoparticles with nearly homogeneous distribution was characterized structurally with TEM, XRD, XPS and elemental mapping. The Pd@p(SPA) was used for the catalytic hydrolysis reaction of ethylenediamine bisborane (EDAB). A series of reactions were completed by changing parameters like the catalyst amount, EDAB concentration and temperature. The Pd@p(SPA) was determined to have turnover frequency (TOF) value of 354 mol H2.molPd(0)−1.h−1 at 30 °C for the hydrolysis reaction of EDAB. The activation parameters for the reaction were Ea = 67.79 kJmol-1, ΔH# = 65.15 kJmol-1 and ΔS# = -103.26 Jmol−1K−1. Pd@p(SPA) also had excellent reusability for the hydrolysis of EDAB. Even after ten reaction cycles, Pd@p(SPA) had 87% catalytic activity. At the same time after four months of storage in an aqueous medium, Pd@p(SPA) still had 100% catalytic activity.

The aim of this research was to design an electrochemical immunosensor for determination of tumour necrosis factor receptor-associated protein-1 TRAP1 antigen, a heat shock protein linked to tumour necrosis factor. The indium-tin oxide covered polyethylene terephthalate ITO-PET electrode surface was cleaned and was prepared for the introduction of hydroxyl groups on its surface by using NH$_{4}$ OH/H$_{2}$O$_{2}$/H$_{2}$O. As a silanization agent for covalent attachment of anti-TRAP1 on the surface of the ITO working electrode, 3-glycidoxypropyltrimethoxysilane 3-GOPS was used. Cyclic voltammetry CV and electrochemical impedance spectroscopy EIS were used to characterize the immobilization steps. A variety of parameters, 3-GOPS and anti-TRAP1 concentrations, and anti-TRAP1 and TRAP1 incubation durations were optimized. After determining the optimum conditions, characterization studies such asrepeatability,reproducibility,regeneration,squarewavevoltammetry,andsinglefrequencyimpedancewereperformed. The electrochemical immunosensor has presented an extremely wide determination range for TRAP1 from 0.1 pg/mL to 100 pg/mL.

In this study, Ru(0) nanoparticles supported in 2-hydroxyethyl starch-p-(2-Acrylamido-2-methyl-1-propanesulfonic acid) interpenetrating polymeric network (HES-p(AMPS) IPN) were synthesized as hydrogel networks containing hydroxyethyl starch, which is a natural polymer with oxygen donor atoms. The structure and morphology of the prepared HES-p(AMPS) IPN hydrogel and Ru@HES-p(AMPS) IPN catalyst were characterized using Fourier transform infrared spectroscopy (FT-IR), scanning electron microscope (SEM), X-ray diffraction (XRD), and transmission electron microscope (TEM). Ru@HES-p(AMPS) IPN was used as catalyst for hydrogen production from the hydrolysis of ethylenediamine bisborane (EDAB). The activation parameters for the hydrolysis reaction of EDAB catalyzed by Ru@HES-p(AMPS) IPN were calculated as Ea = 38.92 kJ mol−1, ΔH# = 36.28 kJ mol−1, and ΔS# = −111.85 J mol−1 K−1, respectively. The TOF for the Ru@HES-p(AMPS) IPN catalyst was 2.253 mol H2 (mol Ru(0) min)−1. It was determined that Ru@HES-p(AMPS) IPN, a reusable catalyst, still had 81.5% catalytic activity after the 5th use.

Bacteria, viruses, and parasites are harmful microorganisms that cause infectious diseases. Early detection of diseases is critical to prevent disease transmission and provide epidemic preparedness, as these can cause widespread deaths and public health crises, particularly in resource-limited countries. Lateral flow assay (LFA) systems are simple-to-use, disposable, inexpensive diagnostic devices to test biomarkers in blood and urine samples. Thus, LFA has recently received significant attention, especially during the pandemic. Here, first of all, the design principles and working mechanisms of existing LFA methods are examined. Then, current LFA implementation strategies are presented for communicable disease diagnoses, including COVID-19, zika and dengue, HIV, hepatitis, influenza, malaria, and other pathogens. Furthermore, this review focuses on an overview of current problems and accessible solutions in detecting infectious agents and diseases by LFA, focusing on increasing sensitivity with various detection methods. In addition, future trends in LFA-based diagnostics are envisioned.

Leptin is a peptide hormone produced in adipose tissue that works as an antiobesity hormone by balancing energy intake and expenditure. We aimed to develop an ultrasensitive electrochemical immunosensor based on a novel immobilization technique for the early detection of leptin-related diseases in this work. Although several methods for immobilizing antibodies to the biosensor recognition element are known, it is necessary to utilize novel, cost-effective, and less complicated immobilization procedures. When compared with currently utilized immobilization techniques for leptin measurement, this novel method is more efficient, easy to prepare, and sensitive, with a broad detection range. Indium tin oxide-coated polyethylene terephthalate (ITO-PET) sheets were used as the working electrode. ITO-PET sheets were modified using cyanogen bromide (CNBr) to immobilize the anti-leptin antibody through covalent interactions. Each stage of the proposed biosensors was characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) methods, and extensive characterization studies were carried out. The designed biosensor has a wide linear detection range (0.05–100 pg/mL), low limits of detection (LOD) (0.0086 pg/mL) and quantification (LOQ) (0.0287 pg/mL). It was concluded that although it is disposable, the ITO-PET working electrode retains its activity even in repeated studies. In addition, the new immobilization procedure provided by CNBr for the designed biosensor fabrication can be effectively used in other biosensing applications. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-03096-w.

An early on time detection of breast cancer significantly affects the treatment process and outcome. Herein, a new label-free impedimetric biosensor is developed to determine the lowest change in the level of calreticulin (CALR), which is a new biomarker of breast carcinoma. The proposed immunosensor is fabricated by using reduced graphene oxide/amino substituted polypyrrole polymer (rGO-PPyNH

Nowadays, breast cancer is one of the riskiest and common cancer types. The early diagnosis of breast cancer is very important just as in other cancer types. Recent investigations demonstrate the b...

A label-free impedimetric immunosensor based on a conjugated poly(pyrrole N-hydroxy succinimide) (PPyr-NHS) polymer covered disposable indium tin oxide electrode (ITO) was fabricated for the ultrasensitive determination of interleukin 6 (IL 6) antigen. In this sensing platform, the PPyr-NHS polymer, which carried a number of succinimide groups on its end side, was used as a matrix material for the first time. This synthesized polymer had excellent biocompatibility and good electrical conductivity. In addition, the utilization of this material as a matrix material provided direct immobilization of the IL 6 receptor (IL6R), which was employed as a biorecognition element. The preparation of the working electrode, the successful attachment of IL 6R and specific interaction between IL6R and IL 6 antigen were confirmed by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) techniques. Moreover, the working electrode surface was characterized by Scanning electron microscopy (SEM), Atomic force microscopy (AFM), Fourier transform infrared spectroscopy (FTIR) and Raman spectroscopy during its construction. An impedimetric method was employed for the quantification of IL 6 antigen. Under optimized experimental conditions, the designed sensing platform could detect IL 6 antigen with a wide detection range from 0.03 pg mL−1 to 22.5 pg mL−1 with a relatively low detection limit of 10.2 fg mL−1. In addition, the developed biosensor had outstanding sensitivity and specificity, good repeatability and reproducibility, high stability and reusability. Additionally, the designed sensing tool was successfully used in human serum samples. Consequently, the suggested immunosensor was clinically useful in the early detection of prostate cancer by direct determination of the serum IL 6 antigen level after simple dilution.

Biosensor technology has been extensively investigated for determination of several analytes in clinical diagnosis, food analysis and environmental monitoring. A biosensor includes three pa...

Background: The determination of drugs in pharmaceutical formulations and human biologic fluids is important for pharmaceutical and medical sciences. Successful analysis requires low sensitivity, high selectivity and minimum interference effects. Current analytical methods can detect drugs at very low levels but these methods require long sample preparation steps, extraction prior to analysis, highly trained technical staff and high-cost instruments. Biosensors offer several advantages such as short analysis time, high sensitivity, real-time analysis, low-cost instruments, and short pretreatment steps over traditional techniques. Biosensors allow quantification not only of the active component in pharmaceutical formulations, but also the degradation products and metabolites in biological fluids. The present review gives comprehensive information on the application of biosensors for drug discovery and analysis. Moreover, this review focuses on the fabrication of these biosensors. Methods: Biosensors can be classified as the utilized bioreceptor and the signal transduction mechanism. The classification based on signal transductions includes electrochemical optical, thermal or acoustic. Electrochemical and optic transducers are mostly utilized transducers used for drug analysis. There are many biological recognition elements, such as enzymes, antibodies, cells that have been used in fabricating of biosensors. Aptamers and antibodies are the most widely used recognition elements for the screening of the drugs. Electrochemical sensors and biosensors have several advantages such as low detection limits, a wide linear response range, good stability and reproducibility. Optical biosensors have several advantages such as direct, real-time and label-free detection of many biological and chemical substances, high specificity, sensitivity, small size and low cost. Modified electrodes enhance sensitivity of the electrodes to develop a new biosensor with desired features. Chemically modified electrodes have gained attention in drug analysis owing to low background current, wide potential window range, simple surface renewal, low detection limit and low cost. Modified electrodes produced by modifying of a solid surface electrode via different materials (carbonaceous materials, metal nanoparticles, polymer, biomolecules) immobilization. Recent advances in nanotechnology offer opportunities to design and construct biosensors. Unique features of nanomaterials provide many advantages in the fabrication of biosensors. Nanomaterials have controllable chemical structures, large surface to volume ratios, functional groups on their surface. To develop proteininorganic hybrid nanomaterials, four preparation methods have been used. These methods are immobilization, conjugation, crosslinking and self-assembly. In the present manuscript, applications of different biosensors, fabricated by using several materials, for drug analysis are reviewed. The biosensing strategies are investigated and discussed in detail. Results: Several analytical techniques such as chromatography, spectroscopy, radiometry, immunoassays and electrochemistry have been used for drug analysis and quantification. Methods based on chromatography require timeconsuming procedure, long sample-preparation steps, expensive instruments and trained staff. Compared to chromatographic methods, immunoassays have simple protocols and lower cost. Electrochemical measurements have many advantages over traditional chemical analyses and give information about drug quantity, metabolic fate of drugs, and pharmacological activity. Moreover, the electroanalytical methods are useful to determine drugs sensitively and selectivity. Additionally, these methods decrease analysis cost and require low-cost instruments and simple sample pretreatment steps. Conclusion: In recent years, drug analyses are performed using traditional techniques. These techniques have a good detection limit, but they have some limitations such as long analysis time, expensive device and experienced personnel requirement. Increased demand for practical and low-cost analytical techniques biosensor has gained interest for drug determinations in medical sciences. Biosensors are unique and successful devices when compared to traditional techniques. For drug determination, different electrode modification materials and different biorecognition elements are used for biosensor construction. Several biosensor construction strategies have been developed to enhance the biosensor performance. With the considerable progress in electrode surface modification, promotes the selectivity of the biosensor, decreases the production cost and provides miniaturization. In the next years, advances in technology will provide low cost, sensitive, selective biosensors for drug analysis in drug formulations and biological samples.

In this study, a sensitive immunosensor was developed for the first time for p21-activated kinase 2 (PAK 2) detection. In the design of the immunosensor, 3-glycidoxypropyltrimethoxysilane (GPTMS) was utilized as an ITO electrode modification material for anti-PAK 2 antibody immobilization. This molecule had epoxy group, which was reactive to amino groups of antibodies. Anti-PAK 2 antibodies were also used as biomolecules for sensitive interaction for PAK 2 antigen. In the presence of PAK 2 antigens, high impedance signal was observed when the process followed by using Electrochemical Impedance Spectroscopy technique (EIS). Apart from EIS technique, Cyclic Voltammetry (CV), Square Wave Voltammetry (SWV) and Single Frequency Impedance (SFI) techniques were utilized. Microscopic surface characterizations of immunosensor fabrication steps were performed by using Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). The immunosensor exhibited good sensitivity and selectivity for PAK 2 antigen detection. A linear calibration curve between EIS response and PAK 2 concentration was obtained in the range of 0.005–0.075 pg/mL with the detection limit of 1.5 fg/mL. It had good repeatability, excellent reproducibility and high stability. Additionally, this immunosensor can be reused by simple application protocol. Furthermore, it had good recovery for PAK 2 antigen detection in human serum samples. The good recovery illustrated that the developed immunosensor was a promising tool for PAK 2 detection in practical applications.

An impedance sensing platform-combined conducting nanocomposite layer was fabricated to develop an effective and rapid method for detection of coronavirus infection (COVID-19) specific spike receptor binding domain (RBD) protein, a precious antigen marker of COVID-19 disease. Coronavirus infection has spread globally and swiftly with major impacts on health, economy, and quality of life of communities. Fast and reliable detection of COVID-19 is a very significant issue for the effective treatment of this bad illness. For this aim, first, an Epoxy functional group-substituted thiophene monomer was synthesized and electrodeposited on a single-use indium tin oxide (ITO) platform in the presence of acetylene black by employing a cyclic voltammetry technique; thus, a conducting nanocomposite (C-NC) layer with high conductivity was obtained. This composite was electrodeposited for the first time on the ITO surface to generate a facile and cost-effective impedimetric biosensor. In addition, this composite provided proper attachment points for antibody binding and also supported the biosensor construction. The immuno-specific biointeractions between anti-RBD and RBD proteins hampered the electron transfer between the ITO substrate surface and electrolyte, and this reaction caused variations in impedance signals, and these signals were proportional to the immobilized RBD antigen amounts. The as-prepared immunosensor showed a wide linear dynamic range (0.0012-120 pg/mL), an ultra-low detection limit of 0.58 fg/mL with added superiorities of great selectivity, suitable repeatability, multiple reusability, and excellent reproducibility.

This study presents a disposable, novel, and sensitive biosensing system to determine adiponectin, an obesity biomarker, in real human serum. The graphite paper (GP) working electrode is a new material for impedimetric biosensors. In the literature, there is no study in which this electrode is used in impedance-based biosensors for adiponectin detection. Sensitive and useful techniques, such as electrochemical impedance spectroscopy and cyclic voltammetry, were utilized for investigation of the modification of the GP electrode surface and optimization and characterization of the constructed biosensor. The single frequency impedance technique was used to study the interactions between antiadiponectin and adiponectin. The morphology of the electrode surface for each immobilization step was examined with scanning electron microscopy. All experimental parameters were optimized to fabricate a rapid and sensitive biosensing system. The designed biosensor presents excellent performance with a wide detection range (0.05-25 pg mL(-1)) and a low limit of detection (0.0033 pg mL(-1)) for adiponectin determination. Also, it has been demonstrated that the biosensor sensitively allows for the detection of adiponectin in human serum. The affinity of the designed immunosensor toward other proteins and components was examined in the presence of the target protein (adiponectin), leptin (100 pg mL(-1) creatine kinase (50 pg mL(-1)), parathyroid hormone (50 pg mL(-1)), and D-glucose (0.5 M). The selectivity of the adiponectin biosensor resulted in high capacity to neglect the interference effect. The constructed biosensor showed good linearity, long-term storage life (10 weeks), high reusability (18 times regenerability), and high ability to detect adiponectin concentrations at picogram levels.

Molecularly imprinted polymer (MIP)-based biosensors have successfully been utilized for sensitive and selective biosensing of different analytes because of imprinted molecular cavities present on the transducer surface. These cavities perfectly match the target analytes and provide the recognition event. After recognition, analytical signals are measured, and these signals determine the target analyte concentration. For construction of MIP-based biosensors, several transduction platforms are utilized and, different applications have been performed by using these biosensing systems. The present chapter summarizes the MIP-based biosensors and evaluates the performance and analytical features of the developed biosensors.

In recent years, advances in immunosensor device fabrication have significantly expanded the use of this technology in a broad range of applications including clinical diagnosis, food analysis, quality control, environmental studies and industrial monitoring. The most important aspect in fabrication is to obtain a design that provides a low detection limit. The utilization of nanomaterials as a label, catalyst and biosensing transducer is, perhaps, the most popular approach in ultrasensitive devices. This chapter reviews recent advances in immunosensor fabrication and summarizes the most recent studies. Strategies employed to significantly improve sensitivity and specificity of immunosensor technology and the advantages and limitations thereof are explored.

The use of quantum dot-based electrochemical immunosensors in a wide perspective from disease markers to agriculture has gained importance in recent years. Due to this importance, many signal amplification strategies have been developed to increase the sensitivity of QD-based immunosensors. As a result of these strategies, immunosensors with high selectivity and sensitivity that are capable of determining many different target molecules with a wide range of determination and with remarkable analytical characteristics have emerged. In this chapter, the properties of QD-based immunosensors are summarized and discussed from a close perspective to some QD-based immunosensors developed in recent years.

In this study, we developed an electrochemical-based single-use neurobiosensor based on multiwalled carbon nanotube (MWCNT)-gold nanoparticle (AuNP) nanocomposite doped, 11-amino-1-undecanethiol (11-AUT)-modified polyethylene terephthalate coated indium tin oxide (ITO-PET) electrodes. This electrode was used for the sensitive determination of DJ-1, a protein responsible for mitochondrial dysfunction in Parkinson's disease (PD) with the task of eliminating oxidative stress. The design strategy and analytical studies for the neurobiosensor were monitored with electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and single frequency impedance (SFI) techniques. The selective determination range for DJ-1 of the developed neurobiosensor system is 4.7–4700 fg mL−1 in accordance with the charge transfer resistance (Rct) associated with a limit of detection of 0.5 fg mL−1. Since changes in the expression of DJ-1 protein is particularly important in cerebrospinal fluid (CSF) and saliva, the ability of the developed neurobiosensor system to detect the DJ-1 protein in these media was tested by the standard addition method. The statistical results show that the biosensor decorated with MWCNT-AuNP-AUT may be recommended for the selective determination of DJ-1 protein.

This study presents a novel, single-use electrochemical biosensor for the leptin biomarker, which may have potential use for early diagnosis of childhood obesity. The graphite paper working electrode was used for the first time in impedimetric biosensors. All immobilization procedure, investigation of the optimal parameters and characterization of biosensors were followed and evaluated using Electrochemical Impedance Spectroscopy (EIS) and Cyclic Voltammetry (CV). The Scanning Electron Microscope (SEM) was utilized to visualize the morphology of the electrode surface during the immobilization steps of the immunosensor. Moreover, the characterization of the interactions between anti-leptin and leptin was investigated by using Single Frequency Technique (SFI). The applicability of the designed biosensor for real serum samples was tested for clinical use. It was observed that the biosensor allows high sensitivity in the analyte detection (leptin) in real serum samples. Moreover, it was suggested that the developed biosensor presents advantages such as long shelf life (5% loss of activity after 8 weeks and 60% loss after 10 weeks), ability to determine analyte concentrations at picogram level (0.2 pg mL−1 -20 pg mL−1), low limit of detection (0.00813 pg mL− 1), reproducibility, reusability (12 times) and high sensitivity.

Herein, a new strategy for the fabrication of a sensitive immunosensor capable of determination of CC Chemokine receptor 4 (CCR4) in complex serum samples is developed through the polymer modification on the disposable indium tin oxide electrode. Anti-CCR4 antibodies, which are utilized as sensing biomolecules, are covalently attached on the succinimide groups of polypyrrole polymer (PPyr-CSsg). The constructed immunosensor illustrates promising performances for the quantification of CCR4 antigen, with a linear detection range of 0.024-12 pg mL-1 and a low detection limit of 7.3 fg mL-1 , calculated at a signal-to-noise ratio of 3. In addition, the impedimetric immunosensor displays a very successful analytical performance in terms of sensitivity, selectivity, repeatability, reproducibility, and long-term stability as well as successful applicability for the accurate quantification of CCR4 in human serum samples. The constructed immunosensor is successfully used for quantification of CCR4 antigen in human serums. In addition, the immunosensor displays only 27.54% loss in its initial signal after nine weeks storage at 4 °C. Moreover, the fabricated immunosensor is economical, highly sensitive, and selective for CCR4 antigen detection, and suitable for potential application in clinical diagnosis.

A new flexible biosensor based on conjugated polymer functionalized indium tin oxide (ITO) sheet was fabricated for Receptor for Activated C Kinase 1 (RACK 1) determination. Poly(3-thiophene acetic acid) (P(Thi-Ac)) was used as an immobilization matrix for construction of RACK 1 immunosensor. This polymer had a great number of carboxyl groups on its end site and these carboxyl ends provided anchoring points to the anti-RACK 1 antibodies. Anti-RACK 1 antibodies were covalently attached on the ITO electrode and recognized the RACK 1 antigens. Electrochemical characterizations were made by employing electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) techniques. Additionally, single frequency impedance method (SFI) was applied to follow the specific biointeraction between antibody and antigen. As a result of specific biointeraction, the designed immunosensor exhibited a wide linear detection range between 0.01 pg/mL and 2 pg/mL RACK 1 with a detection limit of 3.1 fg/mL. Scanning electron microscopy and atomic force microscopy analyses were employed for electrode surface morphology investigation. The designed RACK 1 biosensor had good repeatability (5.73 %, RSD), excellent reproducibility (2.5 %, RSD), long storage-stability and reusable property. In addition, the fabricated RACK 1 biosensor was applied to determine RACK 1 concentration in human serums and the recovery was ranging from 98.79%-100.22%. This work illustrated a new tool to construct a sensitive and low-cost disposable biosensor for applications in clinical monitoring.

A novel label-free impedimetric immunosensor was fabricated for rapid, selective and sensitive detection and quantification of Cadherin-like protein 22 (CDH22), a cancer marker, in human serum by using easy and quickly prepared disposable ITO immunoelectrode. cancer marker, in human serum by using easy and quickly prepared disposable ITO immunoelectrode. The biosensing approach implied the use of ITO electrode coated with poly(phosphazene) polymer including benzaldehyde groups attached with CDH22 antibody and CDH22 antigens. Benzaldehyde side groups containing poly(phosphazene) film coated disposable ITO electrode were utilized as an immunosensing platform and anti-CDH22 antibodies bound to aldehyde groups of benzaldehyde substituted poly(phosphazene) (P-PHP) covalently. The immunosensor modification steps and affinity interaction between anti-CDH22 antibodies and CDH22 antigens were observed by EIS and CV in the presence of the redox couple. Furthermore, antibody immobilization was followed via FTIR and Raman spectroscopy. The morphological analyses of the suggested immunosensor during the fabrication steps were carried out with SEM and AFM monitoring. All the experimental parameters affecting the construction of the immunoelectrodes were optimized. The fabricated immunosensor exhibited an excellent working performance with a wide detection linear range (0.015–2.9 pg/mL) and low limit of detection (4.4 fg/mL). Moreover, the proposed immunosensor had great reproducibility, repeatability and long-term stability. Additionally, the fabricated immunosensor was successfully used in the quantification of CDH22 in human serum without any pretreatment.