Five years of advances in electrochemical analysis of protein biomarkers in lung cancer: a systematic review.

Lung cancer is the leading cause of cancer death in both men and women. It represents a public health problem that must be addressed through the early detection of specific biomarkers and effective treatment. To address this critical issue, it is imperative to implement effective methodologies for specific biomarker detection of lung cancer in real clinical samples. Electrochemical methods, including microfluidic devices and biosensors, can obtain robust results that reduce time, cost, and assay complexity. This comprehensive review will explore specific studies, methodologies, and detection limits and contribute to the depth of the discussion, making it a valuable resource for researchers and clinicians interested in lung cancer diagnosis.

Electrochemical microfluidic immunosensor with graphene-decorated gold nanoporous for T-2 mycotoxin detection.

T-2 is one of the most potent cytotoxic food-borne mycotoxins. In this work, we have developed and characterized an electrochemical microfluidic immunosensor for T-2 toxin quantification in wheat germ samples. T-2 toxin detection was carried out using a competitive immunoassay method based on monoclonal anti-T-2 antibodies immobilized on the poly(methyl methacrylate) (PMMA) microfluidic central channel. The platinum wire working electrode at the end of the channel was in situ modified by a single-step electrodeposition procedure with reduced graphene oxide (rGO)-nanoporous gold (NPG). T-2 toxin in the sample was allowed to compete with T-2-horseradish peroxidase (HRP) conjugated for the specific recognizing sites of immobilized anti-T-2 monoclonal antibodies. The HRP, in the presence of hydrogen peroxide (H2O2), catalyzes the oxidation of 4-tert-butylcatechol (4-TBC), whose back electrochemical reduction was detected on the nanostructured electrode at -0.15 V. Thus, at low T-2 concentrations in the sample, more enzymatically conjugated T-2 will bind to the capture antibodies, and, therefore, a higher current is expected. The detection limits found for electrochemical immunosensor, and commercial ELISA procedure were 0.10 µg kg-1 and 10 µg kg-1, and the intra- and inter-assay coefficients of variation were below 5.35% and 6.87%, respectively. Finally, our microfluidic immunosensor to T-2 toxin will significantly contribute to faster, direct, and secure in situ analysis in agricultural samples.

Advances in Nanomaterials and Composites Based on Mesoporous Materials as Antimicrobial Agents: Relevant Applications in Human Health.

Nanotechnology has emerged as a cornerstone in contemporary research, marked by the advent of advanced technologies aimed at nanoengineering materials with diverse applications, particularly to address challenges in human health. Among these challenges, antimicrobial resistance (AMR) has risen as a significant and pressing threat to public health, creating obstacles in preventing and treating persistent diseases. Despite efforts in recent decades to combat AMR, global trends indicate an ongoing and concerning increase in AMR. The primary contributors to the escalation of AMR are the misuse and overuse of various antimicrobial agents in healthcare settings. This has led to severe consequences not only in terms of compromised treatment outcomes but also in terms of substantial financial burdens. The economic impact of AMR is reflected in skyrocketing healthcare costs attributed to heightened hospital admissions and increased drug usage. To address this critical issue, it is imperative to implement effective strategies for antimicrobial therapies. This comprehensive review will explore the latest scientific breakthroughs within the metal-organic frameworks and the use of mesoporous metallic oxide derivates as antimicrobial agents. We will explore their biomedical applications in human health, shedding light on promising avenues for combating AMR. Finally, we will conclude the current state of research and offer perspectives on the future development of these nanomaterials in the ongoing battle against AMR.

Microfluidic Platform Integrated with Carbon Nanofibers-Decorated Gold Nanoporous Sensing Device for Serum PSA Quantification.

Prostate cancer is a disease with a high incidence and mortality rate in men worldwide. Serum prostate-specific antigens (PSA) are the main circulating biomarker for this disease in clinical practices. In this work, we present a portable and reusable microfluidic device for PSA quantification. This device comprises a polymethyl methacrylate microfluidic platform coupled with electrochemical detection. The platinum working microelectrode was positioned in the outflow region of the microchannel and was modified with carbon nanofibers (CNF)-decorated gold nanoporous (GNP) structures by the dynamic hydrogen bubble template method, through the simultaneous electrodeposition of metal precursors in the presence of CNF. CNF/GNP structures exhibit attractive properties, such as a large surface to volume ratio, which increases the antibody's immobilization capacity and the electroactive area. CNFs/GNP structures were characterized by scanning electron microscopy, energy dispersive spectrometry, and cyclic voltammetry. Anti-PSA antibodies and HRP were employed for the immune-electrochemical reaction. The detection limit for the device was 5 pg mL-1, with a linear range from 0.01 to 50 ng mL-1. The coefficients of variation within and between assays were lower than 4.40%, and 6.15%, respectively. Additionally, its clinical performance was tested in serum from 30 prostate cancer patients. This novel device was a sensitive, selective, portable, and reusable tool for the serological diagnosis and monitoring of prostate cancer.

Highly sensitive and selective nanostructured microbiosensors for glucose and lactate simultaneous measurements in blood serum and in vivo in brain tissue.

Highly sensitive and selective nanostructured lactate and glucose microbiosensors for their in vivo simultaneous determination in rat brain were developed based on carbon fiber microelectrodes (CFM) modified with nanoporous gold (NPG) using the Dynamic Hydrogen Bubble Template (DHBT) method. Electrodeposition of platinum nanoparticles (PtNP) onto the NPG film enhances the sensitivity and the electrocatalytic properties towards H2O2 detection. The nanostructured microelectrode platform was modified by glucose oxidase (GOx) and lactate oxidase (LOx) enzyme immobilization. High selective measurements were achieved by covering with a perm-selective layer of electropolymerized m-phenylenediamine, deposition of a Nafion® film and by using a null sensor. The morphological characteristics and electroanalytical performance of the microbiosensors were assessed, by scanning electron microscopy and electrochemical techniques, respectively. The PtNP/NPG/CFM shows a high sensitivity to H2O2 (5.96 A M-1 cm-2) at 0.36 V vs. Ag/AgCl, with a linear range from 0.2 to 200 µM, and an LOD of 10 nM. The microbiosensors were applied to the simultaneous determination of lactate and glucose in blood serum samples. Moreover, the basal extracellular concentrations of lactate and glucose were measured in vivo in four different rat brain structures. These results support the potential of the microbiosensor to be used as a valuable tool to investigate brain neurochemicals in vivo.

Diagnostic Methods for Non-Falciparum Malaria.

Malaria is a serious public health problem that affects mostly the poorest countries in the world, killing more than 400,000 people per year, mainly children under 5 years old. Among the control and prevention strategies, the differential diagnosis of the Plasmodium-infecting species is an important factor for selecting a treatment and, consequently, for preventing the spread of the disease. One of the main difficulties for the detection of a specific Plasmodium sp is that most of the existing methods for malaria diagnosis focus on detecting P. falciparum. Thus, in many cases, the diagnostic methods neglect the other non-falciparum species and underestimate their prevalence and severity. Traditional methods for diagnosing malaria may present low specificity or sensitivity to non-falciparum spp. Therefore, there is high demand for new alternative methods able to differentiate Plasmodium species in a faster, cheaper and easier manner to execute. This review details the classical procedures and new perspectives of diagnostic methods for malaria non-falciparum differential detection and the possibilities of their application in different circumstances.

Fabrication of dendritic nanoporous gold via a two-step amperometric approach: Application for electrochemical detection of methyl parathion in river water samples.

In this work, nanoporous gold (NPG) was prepared according to three different approaches, such as (i) anodization-electrochemical reduction (A-ECR, NPGA), (ii) dynamic hydrogen bubble template (DHBT, NPGB), and (iii) the combination of both methods (NPGA+B). Field-emission scanning electron microscopy (FE-SEM) and cyclic voltammetry (CV) were used to investigate the structural morphology and the electrochemical behavior of the fabricated materials. The NPGA+B electrode showed a large amount of surface defects and/or edges, greater electrochemical surface area (2.5 cm2), and increased roughness factor (35.4). Such outstanding features of the NPGA+B platform were demonstrated by the sensitive detection of methyl parathion (MP) in river water samples. CV results indicated nearly 25-fold, 6-fold, and 2.5-fold higher sensitivity for NPGA+B compared to that of bare Au, NPGA, and NPGB, respectively. Differential pulse voltammetry (DPV) results show a linear behavior in the MP concentration range of 5-50 ng mL-1 with a limit of detection (LOD) of 0.6 ng mL-1 and limit of quantification (LOQ) of 2.0 ng mL-1. Besides, the NPGA+B sensor also revealed excellent selectivity towards MP detection in the presence of other interfering molecules or ions, reproducibility, and repeatability.

Sandwich-Type Electrochemical Paper-Based Immunosensor for Claudin 7 and CD81 Dual Determination on Extracellular Vesicles from Breast Cancer Patients.

This study is focused on identifying novel epithelial markers in circulating extracellular vesicles (EVs) through the development of a dual sandwich-type electrochemical paper-based immunosensor for Claudin 7 and CD81 determination, as well as its validation in breast cancer (BC) patients. This immunosensor allows for rapid, sensitive, and label-free detection of these two relevant BC biomarkers. Under optimum conditions, the limit of detection for Claudin 7 was 0.4 pg mL-1, with a wide linear range of 2 to 1000 pg mL-1, while for CD81, the limit of detection was 3 pg mL-1, with a wide linear range of 0.01 to 10 ng mL-1. Finally, we validated Claudin 7 and CD81 determination in EVs from 60 BC patients and 20 healthy volunteers, reporting higher diagnostic accuracy than the one observed with classical diagnostic markers. This analysis provides a low-cost, specific, versatile, and user-friendly strategy as a robust and reliable tool for early BC diagnosis.

Ultrasensitive microfluidic electrochemical immunosensor based on electrodeposited nanoporous gold for SOX-2 determination.

An ultrasensitive and portable microfluidic electrochemical immunosensor for SOX-2 cancer biomarker determination was developed. The selectivity and sensitivity of the sensor were improved by modifying the microfluidic channel. This was accomplished through a physical-chemical treatment to produce a hydrophilic surface, with an increased surface to volume/ratio, where the anti-SOX-2 antibodies can be covalently immobilized. A sputtered gold electrode was used as detector and its surface was activated by using a dynamic hydrogen bubble template method. As a result, a gold nanoporous structure (NPAu) with outstanding properties, like high specific surface area, large pore volume, uniform nanostructure, good conductivity, and excellent electrochemical activity was obtained. SOX-2 present in the sample was bound to the anti-SOX-2 immobilized in the microfluidic channel, and then was labeled with a second antibody marked with horseradish peroxidase (HRP-anti-SOX-2) like a sandwich immunoassay. Finally, an H2O2 + catechol solution was added, and the enzymatic product (quinone) was reduced on the NPAu electrode at +0.1 V (vs. Ag). The current obtained was directly proportional to the SOX-2 concentration in the sample. The detection limit achieved was 30 pg mL-1, and the coefficient of variation was less than 4.75%. Therefore, the microfluidic electrochemical immunosensor is a suitable clinical device for in situ SOX-2 determination in real samples.

One-step enzyme-free dual electrochemical immunosensor for histidine-rich protein 2 determination.

In the present work, we describe a novel one-step enzyme-free dual electrochemical immunosensor for the determination of histidine-rich protein 2 (Ag-PfHRP2), a specific malaria biomarker. A gold electrode (GE) was functionalized with the PfHRP2 antibody (Ab-PfHRP2) using dihexadecyl phosphate (DHP) polymer as an immobilization platform. The Ab-PfHRP2/DHP/GE sensor was characterized by cyclic voltammetry, electrochemical impedance spectroscopy, Fourier-transform infrared spectroscopy, scanning electron microscopy, and atomic force microscopy. The developed immunosensor was employed for indirect Ag-PfHRP2 determination by differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). The linear range was 10-400 ng mL-1 and 10-500 ng mL-1 for EIS and DPV, while the limit of detection was 3.3 ng mL-1 and 2.8 ng mL-1, respectively. The electrochemical immunosensor was successfully applied for Ag-PfHRP2 determination in human serum samples. Its performance was compared with an ELISA test, and good correspondence was achieved. The coefficients of intra- and inter-assay variations were less than 5%. The electrochemical immunosensor is a useful and straightforward tool for in situ malaria biomarker determination.

Electrochemical microfluidic immunosensor based on TES-AuNPs@Fe3O4 and CMK-8 for IgG anti-Toxocara canis determination.

We report a microfluidic immunosensor for the electrochemical determination of IgG antibodies anti-Toxocara canis (IgG anti-T. canis). In order to improve the selectivity and sensitivity of the sensor, core-shell gold-ferric oxide nanoparticles (AuNPs@Fe3O4), and ordered mesoporous carbon (CMK-8) in chitosan (CH) were used. IgG anti-T. canis antibodies detection was carried out using a non-competitive immunoassay, in which excretory secretory antigens from T. canis second-stage larvae (TES) were covalently immobilized on AuNPs@Fe3O4. CMK-8-CH and AuNPs@Fe3O4 were characterized by transmission electron microscopy, scanning electron microscopy, energy dispersive spectrometry, cyclic voltammetry, electrochemical impedance spectroscopy, and N2 adsorption-desorption isotherms. Antibodies present in serum samples immunologically reacted with TES, and then were quantified by using a second antibody labeled with horseradish peroxidase (HRP-anti-IgG). HRP catalyzes the reduction from H2O2 to H2O with the subsequent oxidation of catechol (H2Q) to p-benzoquinone (Q). The enzymatic product was detected electrochemically at _100 mV on a modified sputtered gold electrode. The detection limit was 0.10 ng mL-1, and the coefficients of intra- and inter-assay variation were less than 6%, with a total assay time of 20 min. As can be seen, the electrochemical immunosensor is a useful tool for in situ IgG antibodies anti-T. canis determination.

Paper surface modification strategies employing N-SBA-15/polymer composites in bioanalytical sensor design.

In this work, different paper surface modification strategies were compared to obtain an amine functionalized SBA-15 (N-SBA-15) composite for paper-based device development. The synthesized N-SBA-15 was characterized by N2 adsorption-desorption isotherm, and infrared spectroscopy (FTIR), and it was incorporated to different polymer matrices (κ-carrageenan (CA), polyvinyl alcohol (PVA) and polyethylenimine (PEI)) for the development of the composite modified paper-based device. The retention, interactions, and morphology of the obtained composites were investigated by absorbance measurement, FTIR and scanning electron microscopy (SEM), respectively. To demonstrate the applicability of the modified paper-based device, ascorbic acid (AA) quantification was carried out. Horseradish peroxidase (HRP) was immobilized onto the modified paper surface. HRP in the presence of H2O2 catalyzes the oxidation of 10-acetyl-3,7-dyhidroxyphenoxazine (ADHP) to highly fluorescent resorufin, which was measured by LIF detector. Thus, when AA was added to the solution, it decreases the relative fluorescence signal proportionally to the AA concentration. The linear range from 50 nmol L-1 to 1500 nmol L-1 and a detection limit of 15 nmol L-1 were obtained for AA quantitation. The obtained results allowed us to conclude that N-SBA-15/PEI composite could be considered an excellent choice for the paper-based device modification procedure due to its inherent simplicity, low cost, and sensitivity.

Development of a nanostructured immunosensor for early and in situ detection of Xanthomonas arboricola in agricultural food production.

We report a microfluidic electrochemical immunosensor for Xanthomonas arboricola (XA) determination, based on the covalently immobilization of monoclonal anti-XA antibody (anti-XA) on a previously amino functionalized SBA-15 in situ synthesized in the central channel of a glass-poly(dimethylsiloxane) microfluidic immunosensor. The synthetized amino-SBA-15 was characterized by N2 adsorption-desorption isotherm, scanning electron microscopy and infrared spectroscopy. XA was detected by a direct sandwich immunoassay through an alkaline phosphatase (AP) enzyme-labeled anti-XA conjugate. Later, the substrate p-aminophenyl phosphate was converted to p-aminophenol by AP. The enzymatic product was detected at +100mV on a sputtered gold electrode. The measured current was directly proportional to the level of XA in walnut trees samples. The linear range was from 5 × 102 to 1 × 104CFUmL-1. The detection limit was 1.5 × 102CFUmL-1, and the within- and between-assay coefficients of variation were below 5%. Microfluidic immunosensor is a very promising tool for the early and in situ diagnosis of XA in walnuts avoiding serious economic losses.

Microfluidic immunosensor based on mesoporous silica platform and CMK-3/poly-acrylamide-co-methacrylate of dihydrolipoic acid modified gold electrode for cancer biomarker detection.

We report a hybrid glass-poly (dimethylsiloxane) microfluidic immunosensor for epidermal growth factor receptor (EGFR) determination, based on the covalent immobilization of anti-EGFR antibody (anti-EGFR) on amino-functionalized mesoporous silica (AMS) retained in the central channel of a microfluidic device. The synthetized AMS was characterized by N2 adsorption-desorption isotherm, scanning electron microscopy (SEM), energy dispersive spectrometry (EDS) and infrared spectroscopy. The cancer biomarker was quantified in human serum samples by a direct sandwich immunoassay measuring through a horseradish peroxidase-conjugated anti-EGFR. The enzymatic product was detected at -100 mV by amperometry on a sputtering gold electrode, modified with an ordered mesoporous carbon (CMK-3) in a matrix of poly-acrylamide-co-methacrylate of dihydrolipoic acid (poly(AC-co-MDHLA)) through in situ copolymerization. CMK-3/poly(AC-co-MDHLA)/gold was characterized by cyclic voltammetry, EDS and SEM. The measured current was directly proportional to the level of EGFR in human serum samples. The linear range was from 0.01 ng mL-1 to 50 ng mL-1. The detection limit was 3.03 pg mL-1, and the within- and between-assay coefficients of variation were below 5.20%. The microfluidic immunosensor is a very promising device for the diagnosis of several kinds of epithelial origin carcinomas.

Copper nanoparticles applied to the preconcentration and electrochemical determination of ß-adrenergic agonist: an efficient tool for the control of meat production.

A novel method for preconcentration and electrochemical detection of zinterol in bovine urine samples was developed. In order to improve the limit of detection, the surface of a screen-printed carbon electrode was modified with electrodeposited metal copper nanoparticles. The experimental electrodeposition optimization was performed using a central composite design (CCD), involving the variables: precursor concentration, potential and time applied. Copper nanoparticles were characterized by transmission electron microscopy, scanning electron microscopy, cyclic voltammetry, and energy dispersive X-ray spectroscopy. Mesoporous shuttle-like copper oxide nanoparticles were used for the preconcentration step to avoid interferences with many compounds present in the sample matrix. The optimal working conditions for the preconcentration approach were found by means of both two-level fractional factorial and CCD designs. The obtained enhancement factor for a sample volume of 30 mL was 35 fold. The calibration curve showed linearity between 0.5 and 45 ng mL(-1) and the limit of detection was 0.16 ng mL(-1). The intra and inter assay coefficients of variability were below 4% and 5%; respectively.

Food safety control of zeranol through voltammetric immunosensing on Au-Pt bimetallic nanoparticle surfaces.

This study reports an accurate and sensitive strategy for zeranol (ZER) determination in bovine urine samples. ZER is a mycotoxin widely used as a synthetic growth promoter in the livestock production whose residues could present a potential risk for human health. Therefore, its use as an animal feed additive has been banned in most countries. ZER determination was accomplished using an electrochemical system in which bimetallic Au-Pt nanoparticles (Au-PtNPs) were electro-synthesized on a screen-printed carbon electrode (SPCE). The obtained Au-PtNP platform was immunofunctionalized using specific anti-ZER antibodies as a strategy to avoid potential interference. After biorecognition, ZER was directly oxidized and detected by square-wave voltammetry (SWV). The Au-PtNP surface was characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD) and cyclic voltammetry (CV). The limit of detection calculated was 0.01 ng mL(-1) with a wide linear range from 0.03 to 30 ng mL(-1). This method promises to be suitable for ZER quantification in bovine urine samples ensuring food quality and safety, as well as consumer's health.

Ultra sensitive microfluidic immunosensor for determination of clenbuterol in bovine hair samples using electrodeposited gold nanoparticles and magnetic micro particles as bio-affinity platform.

In this article, we report the first integrated microfluidic immunosensor coupled to a screen-printed carbon electrode (SPCE) applied to determination of clenbuterol (CLB) in bovine hair samples. CLB is a member of the ß(2)-agonist drugs which is used in animal production and is banned in Argentine and the European Union. It represents a potential risk and has to be carefully monitored to avoid the illegal use of high amounts of this compound that could result in human food poisoning. In order to perform the CLB detection, the SPCE was modified by gold nanoparticles (AuNPs) electrodeposition. Quantitative determination of CLB was carried out using a competitive indirect immunoassay, method based on the use of anti-CLB antibodies immobilized on magnetic micro particles. The CLB present in bovine hair samples competes immunologically with alkaline phosphatase (AP) enzyme-labeled CLB conjugate for the anti-CLB specific antibodies. Later, p-aminophenyl phosphate was converted to p-aminophenol by AP, and the electroactive product was quantified on AuNPs/SPCE at +0.1 V. The limit of detection for electrochemical method was 0.008 ng mL(-1) and the intra- and inter-assay coefficients of variation were below 6%. This being a veterinary control tool very useful for rapid, sensitive and selective detection of CLB in an "in vitro" technique.