Hand-Operated, Paper-Based Rotational Vertical-Flow Immunosensor for the Impedimetric Detection of α-Fetoprotein.

This study demonstrates a hand-operated, paper-based rotational vertical-flow immunosensor (rotational VFI) platform requiring fewer pipetting steps, designed for the electrochemical detection of α-fetoprotein with multiple and time-sequenced steps. The platform allows users to perform electrochemical measurements without interference from the convective component of fluid motion, which is unfavorable in most techniques. Users can freely transfer-switch-stop fluid flows by manually rotating the paper disk, evidencing the superior flexibility of this sensor compared to other biosensors. Furthermore, the overall assay duration can be considerably shortened to 9 min. The linear range (LR) is determined to be 0.01-500 ng/mL, with a limit of detection (LOD) of 1.65 pg/mL, and the sensitivity can be significantly enhanced simply by switching off the sample stream to ensure detention at the binding zone (for up to 30 min). This additional step can widen the LR to 0.5 pg/mL, with a LOD of 3.54 fg/mL, which is the lowest detectable level ever reported among paper-based sensors. The advantages of the designed rotational VFI qualify it as a suitable alternative to various biosensors.

Resistance-Based Lateral Flow Immunosensor with a NFC-Enabled Smartphone for Rapid Diagnosis of Leptospirosis in Clinical Samples.

Leptospirosis is one of the most life-threatening tropical diseases caused by pathogenic Leptospira. To date, a diagnostic device that offers rapid and sensitive detection of leptospires has been still in demand for proper treatment to reduce the mortality rate. Herein, we create a resistance-based lateral flow immunosensor diagnosis device (R-LFI) that integrates near-field communication (NFC) with a portable smartphone for leptospiral detection in clinical samples. A specific monoclonal antibody against the pathogen was coated on a nitrocellulose membrane (NCM) where the test line was collocated. Two electrodes with a sandwich-like configuration were installed employing a conductive double-sided adhesive tape and connected with a NFC smartphone-based detection system. A half-sandwich immunocomplex formation induced high proton conduction, resulting in a considerable decrement in resistive response. The performance of the R-LFI sensor was evaluated using recombinant LipL32 (rLipL32), Leptospira interrogans, and clinical samples. The R-LFI device exhibited linear responses toward rLipL32 protein in phosphate buffer and L. interrogans-spiked healthy human serum samples within the concentration ranging from 1 to 1000 ng mL-1 (limit of detection (LOD): 0.29 ng mL-1) and from 104 to 106 cell mL-1 (LOD: 4.89 × 103 cell mL-1), respectively. Our R-LFI sensor successfully detected L. interrogans-positive clinical samples as confirmed by polymerase chain reaction (PCR). This platform offers high specificity, selectivity, simplicity, miniscule sample volume, and no labeling element requirement. These desirable features make it particularly suitable for countries where medical facilities and resources are limited.

Voltammetric lipase activity assay based on dilinolein and a modified carbon paste electrode.

In this work, a novel electrochemical assay for characterizing both lipases and lipase inhibitors as well as for the determination of lipase activity is described. It is based on a carbon paste electrode, modified with cobalt(II)phthalocyanine, and multi-walled carbon nanotubes (MWCNTs). As reaction media, a sodium borate buffer was used (0.1 M, pH 9). The measurements were carried out in a batch system using differential pulse voltammetry (DPV) and 1,3-dilinolein as standard substrate. The activity assay showed a linearity for porcine pancreas lipase activity in a range between 20 and 300 U L-1 (per min) with a limit of detection (LOD) of 7 U L-1 and a limit of quantification (LOQ) of 20 U L-1. The kinetic behavior of the lipase reaction was investigated, resulting in a KM value of 0.29 mM. The applicability of the activity assay could be shown by investigating the activity of lipases from Aspergillus oryzae and Candida rugosa, and the results were confirmed by a reference method. The inhibitory effects were characterized with Orlistat.

An electrochemical sensor for the voltammetric determination of artemisinin based on carbon materials and cobalt phthalocyanine.

An electrochemical sensor for the determination of artemisinin has been developed based on a glassy carbon electrode modified with hybrid nanocomposites of cobalt phthalocyanine, graphene nanoplatelets, multi-walled carbon nanotubes and ionic liquids (IL). To improve the sensitivity and selectivity of the sensor, cobalt phthalocyanine (CoPc) was used as an effective redox mediator to promote and catalyze the artemisinin reduction. Furthermore, the graphene nanoplatelets and multi-walled carbon nanotubes were used as excellent conducting supporting materials to improve the sensitivity of the electrochemical sensor. Moreover, IL with a surface charge was also employed to prevent aggregation of the graphene nanoplatelets and multi-walled carbon nanotubes. The analytical signal was generated from the reduction of Co(III)Pc generated by artemisinin. The proposed electrochemical sensor was applied to the detection of artemisinin using differential pulse voltammetry and provided a signal with wide linearity ranging from 1.5-60 µM and 60-600 µM and a detection limit of 0.70 µM (3SD/m). Furthermore, the proposed sensor displayed good repeatability and reproducibility of 2.9-3.0 and 3.1-4.4% RSD, respectively. Applications of the sensor to drug and plant samples demonstrated accuracy in a range of 105-116% recoveries. In addition, the results were in good agreement with those obtained from the HPLC method as a reference technique. Thus, the proposed electrochemical sensor provides a new alternative platform for sensitive and selective determination of artemisinin in the analysis of pharmaceuticals with good precision and accuracy.

Enzymatic electrochemical biosensor for glyphosate detection based on acid phosphatase inhibition.

A novel enzymatic electrochemical biosensor was fabricated for the indirect detection of glyphosate-based acid phosphatase inhibition. The biosensor was constructed on a screen-printed carbon electrode modified with silver nanoparticles, decorated with electrochemically reduced graphene oxide, and chemically immobilized with acid phosphatase via glutaraldehyde cross-linking. We measured the oxidation current by chronoamperometry. The current arose from the enzymatic reaction of acid phosphatase and the enzyme-substrate disodium phenyl phosphate. The biosensing response is a decrease in signal resulting from inhibition of acid phosphatase in the presence of glyphosate inhibitor. The inhibition of acid phosphatase by glyphosate was investigated as a reversible competitive-type reaction based on the Lineweaver-Burk equation. Computational docking confirmed that glyphosate was the inhibitor bound in the substrate-binding pocket of acid phosphatase and that it was able to inhibit the enzyme efficiently. Additionally, the established method was applied to the selective analysis of glyphosate in actual samples with satisfactory results following a standard method.

Laser engraved microapillary pump paper-based microfluidic device for colorimetric and electrochemical detection of salivary thiocyanate.

A microcapillary grooved paper-based analytical device capable of dual-mode sensing (colorimetric and electrochemical detection) was demonstrated for analysis of viscous samples (e.g., human saliva). Herein, a hollow capillary channel was constructed via laser engraved micropatterning functions as a micropump to facilitate viscous fluidic transport, which would otherwise impede analysis on paper devices. Using salivary thiocyanate as a model analyte, the proposed device was found to exhibit a promising sensing ability on paper devices without the need for sample pretreatment or bulky instrumentation, as normally required in conventional methods used for saliva analysis. An extensive linear dynamic range covering detection of salivary thiocyanate for both high and trace level regimes (5 orders of magnitude working range) was collectively achieved using the dual-sensing modes. Under optimal conditions, the limit of detection was 6 µmol L-1 with a RSD of less than 5%. An excellent stability for the µpumpPAD was also observed for over 30 days. Real sample analysis using the proposed device was found to be in line with the standard chromatographic method. Benefitting from simple fabrication and operation, portability, disposability, low sample volume (20 µL), and low cost (< 1 USD), the µpumpPAD is an exceptional alternative tool for the detection of various biomarkers in saliva specimens.

Highly Sensitive Amperometric Detection of Hydrogen Peroxide in Saliva Based on N-Doped Graphene Nanoribbons and MnO2 Modified Carbon Paste Electrodes.

Four different graphene-based nanomaterials (htGO, N-htGO, htGONR, and N-htGONR) were synthesized, characterized, and used as a modifier of carbon paste electrode (CPE) in order to produce a reliable, precise, and highly sensitive non-enzymatic amperometric hydrogen peroxide sensor for complex matrices. CPE, with their robustness, reliability, and ease of modification, present a convenient starting point for the development of new sensors. Modification of CPE was optimized by systematically changing the type and concentration of materials in the modifier and studying the prepared electrode surface by cyclic voltammetry. N-htGONR in combination with manganese dioxide (1:1 ratio) proved to be the most appropriate material for detection of hydrogen peroxide in pharmaceutical and saliva matrices. The developed sensor exhibited a wide linear range (1.0-300 µM) and an excellent limit of detection (0.08 µM) and reproducibility, as well as high sensitivity and stability. The sensor was successfully applied to real sample analysis, where the recovery values for a commercially obtained pharmaceutical product were between 94.3% and 98.0%. Saliva samples of a user of the pharmaceutical product were also successfully analyzed.

Flow Injection Amperometric Evaluation of Trolox Equivalent Antioxidant Capacity of Chocolates with Different Cocoa Content at a Boron-Doped Diamond Electrode.

RESEARCH BACKGROUND: The objective of this paper is to introduce an instrumentally simple analytical tool for determination of cocoa solid content in chocolates. This electroanalytical method is based on amperometric oxidation of all present antioxidants in chocolates at boron-doped diamond electrode (BDDE) that is integrated in a flow injection analysis (FIA) wall-jet electrode system. EXPERIMENTAL APPROACH: As part of optimisation, thirteen commonly occurring antioxidants were investigated using cyclic voltammetry at the BDDE in 0.1 mol/L phosphate buffer with different methanol (MeOH) content. Working parameters, such as MeOH volume fraction, flow rate and detection potential, were optimised. Principally, the height of the oxidation peak (current response) representing the oxidation of the sum of antioxidants (total antioxidant content; TAC) was expressed as Trolox equivalents. RESULTS AND CONCLUSIONS: For analytical purpose, a linear range from 5 to 100 mg/L described by regression equation and characterised by high correlation coefficient R2=0.9994 was achieved. Obtained high positive correlation between the determined values of Trolox equivalent antioxidant capacity (TEAC) and cocoa mass fractions characterised by correlation coefficient of 0.9187 for eight randomly selected samples (one white, two milk, and five dark chocolates) confirmed that cocoa solids represent the main source of antioxidants (reducing agents). NOVELTY AND SCIENTIFIC CONTRIBUTION: The research demonstrates that TEAC values could be considered as an additional marker of cocoa content in the chocolate analysis to the commonly used theobromine (authenticity of food products). The developed FIA could therefore serve as simple analytical tool in the food quality control.

Wide electrochemical window of screen-printed electrode for determination of rapamycin using ionic liquid/graphene composites.

A disposable screen-printed carbon electrode (SPCE) modified with an ionic liquid/graphene composite (IL/G) exhibits a wider potential window, excellent conductivity, and specific surface area for the improvement in the voltammetric signal of rapamycin detection. The modified composite was characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), and electrochemical impedance spectroscopy (EIS). The electrochemical behavior of rapamycin at the modified SPCE was investigated by cyclic and square wave voltammetry in 60:40 EtOH: 0.1 M LiClO4 at pH 5.0. A high reproducible and well-defined peak with a high peak current were obtained for rapamycin detection at a position potential of + 0.98 V versus Ag/AgCl. Under the optimized conditions, the rapamycin concentration in the range 0.1 to 100 µM (R2 = 0.9986) had a good linear relation with the peak current. The detection limit of this method was 0.03 µM (3SD/slope). The proposed device can selectively detect rapamycin in the presence of commonly interfering compounds. Finally, the proposed method was successfully applied to determine rapamycin in urine and blood samples with excellent recoveries. These devices are disposable and cost-effective and might be used as an alternative tool for detecting rapamycin in biological samples and other biological compounds. Graphical abstract Schematic presentation of wide electrochemical window and disposable screen-printed sensor using ionic liquid/graphene composite for the determination of rapamycin. This composite can enhance the oxidation current and expand the potential for rapamycin detection.

A novel paper-based colorimetry device for the determination of the albumin to creatinine ratio.

A novel paper-based analytical device (PAD) was fabricated and developed for the simple and rapid determination of the albumin to creatinine ratio (ACR) in urine samples. The detection was based on a colorimetric reaction using bromocresol green (BG) in a phosphate buffer (PB) at pH 4 for the determination of albumin (AL) and creatinine (CR). BG changes color from greenish-yellow to bluish-green in the presence of AL and/or CR. Picric acid (PA) in 0.25 M NaOH was used to detect CR, and PA changes color from yellow to orange. Under the optimal conditions, the working range was 10 to 350 mg dL-1 with a detection limit of 7.1 and 5.4 mg dL-1 for AL + CR and CR detection, respectively. The repeatability was evaluated, and the %RSD value was less than 8.23 (n = 10). The ACR was obtained by calculating the AL and CR colorimetric results. Finally, the proposed devices were applied to the determination of AL, CR, and ACR in urine samples. The results obtained by the developed PADs were in good agreement with the standard method and demonstrated the method could reliably measure AL, CR, and ACR. The proposed method provides a low-cost, simple, sensitive, and promising tool for diagnostic identification assay for chronic kidney disease (CKD).

Rapid Electrochemical Method for the Determination of L-DOPA in Extract From the Seeds of Mucuna Prurita.

This work presents the electrochemical behavior of levodopa (L-DOPA), at boron-doped diamond (BDD) electrodes, using cycling voltammetry (CV), in Britton-Robinson (BR) buffer solution, and application of the proposed electrode for the determination of L-DOPA in extracts from the seeds of velvet bean (Mucuna prurita Hook or Mucuna pruriens (L.) DC.). L-DOPA provides a well-defined and single oval-shape oxidation peak at +0.8 V vs. Ag/AgCl (3 M KCl) reference electrode in BR buffer solution at pH 3.0. Experimental parameters, such as pH of supporting electrolyte and square wave voltammetry (SWV) operating parameters (frequency and modulation amplitude) were optimized. The effect of possible interferences was evaluated. Under optimal conditions the detection limit of the developed method was 0.8 µM and the calibration curve for L-DOPA was linear in the range from 2 to 100 µM. The proposed method was successfully applied to the determination of L-DOPA in an extract from the seeds of Mucuna prurita. The obtained result was in good agreement with obtained by photometry with 2,2´-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS). The developed approach can be beneficial for the quantification of L-DOPA using a BDD electrode as up-to-date potential alternative sensor for electroanalytical applications.

Electrochemical determination of riboflavin in pharmaceuticals using unmodified screen printed carbon electrodes.

In this study, we have devised an efficient and rapid approach to detect riboflavin (also known as Vitamin B2 or VB2) utilizing an unaltered screen-printed carbon electrode (SPCE). The unmodified screen-printed electrodes are created within the laboratory, where carbon ink is applied to a ceramic substrate. All experiments pertaining to the investigation of electrochemical behavior and the fine-tuning of crucial experimental parameters were conducted through cyclic voltammetry (CV). For quantitative assessments, square wave voltammetry (SWV) was employed. The findings indicate that unaltered SPCEs exhibit robust current signals during the riboflavin redox reaction. Riboflavin displays a distinct oxidation peak at - 0.136 V (vs. Ag/AgCl, 3.0 M KCl) in a Britton-Robinson buffer solution (BRBS) at pH 2, which was employed for quantification. The electrode demonstrates a broad linear range from 0.05 to 10 µM, boasting a detection limit of 0.03 µM. Repeatability stands at 1.45%, while reproducibility is 6.61%. Testing the influence of common interfering compounds yielded negligible results. The sensor effectively determines riboflavin content in pharmaceutical formulations without any prior treatment. This method presents an economical, modifier-free sensor with exceptional sensitivity and cost-effectiveness, making it suitable for rapid riboflavin quantification.

Paper-based electrochemical immunosensor for the determination of symmetric dimethylarginine.

In this work, we present the development of an immunosensor for the direct, selective, and sensitive determination of symmetric dimethylarginine (SDMA) in urine, in view of the emerging role of this molecule as a biomarker for renal disease. SDMA is almost completely excreted by the kidneys, hence in renal dysfunction, the excretion is decreased, resulting in accumulation in plasma. Reference values for plasma or serum have already been established in small animal practice. Values < 15 µg/dL are considered normal, 15-19 µg/dL are values of concern, and at values > 20 µg/dL kidney disease is likely. The proposed electrochemical paper-based sensing platform uses anti-SDMA antibodies for targeted detection of SDMA. Quantification is related to a decrease in the signal of a redox indicator due to the formation of an immunocomplex that interferes with electron transfer. Square wave voltammetry measurements showed a linear correlation of the peak decline for 50 nM - 1 µM SDMA with a detection limit of 15 nM. The influence of common physiological interferences caused no significant peak reduction, indicating excellent selectivity. The proposed immunosensor was successfully applied for the quantification of SDMA in human urine of healthy individuals. Surveillance of SDMA concentration in urine could prove to be very valuable in the diagnosis or monitoring of renal disease.

Rapid electrochemical lateral flow device for the detection of Δ9-tetrahydrocannabinol.

Cannabis is a plant that is harmful and beneficial because it contains more than 400 bioactive compounds, and the main compounds are Δ9 tetrahydrocannabinol (THC) and cannabidiol (CBD). Currently, cannabis extracts are used in medicine, but the amount of THC as a main psychoactive component is strictly regulated. Therefore, the ability to rapidly and accurately detect THC is important. Herein, we developed a sensitive electrochemical method combining a rapid lateral flow assay (LFA) to detect THC rapidly. An electrochemical LFA device was constructed by attaching a screen-printed electrode inside a lateral-flow device to exploit the remarkable binding of THC to the cannabinoid type 2 (CB2) receptor in the test zone. The ferrocene carboxylic acid attached to the monoclonal THC antibody acts as an electroactive species when it binds to the THC in the sample before it flows continuously to the CB2 receptor region on the electrode. Under optimal conditions, the detection time is within 6 min and the devise shows excellent performance with a detection limit of 1.30 ng/mL. Additionally, the device could be applied to detect THC in hemp extract samples. The results obtained from this sensor are similar to the standard method (HPLC) for detecting THC. Therefore, this proposed device is useful as an alternative device for the on-site determination of THC because it is inexpensive, portable, and exhibits high sensitivity.

A novel immunosensor based on cobalt oxide nanocomposite modified single walled carbon nanohorns for the selective detection of aflatoxin B1.

We developed a novel, sensitive, and selective platform for the specific determination of aflatoxin B1 (AFB1). Single-walled carbon nanohorns decorated by a cobalt oxide composite and gold nanoparticles were created to provide facile electron transfer and improve the sensor's sensitivity. In addition, we attributed the selectivity of the proposed sensor to the specific binding property of the anti-aflatoxin B1 antibody. We clarified the specific interaction of the proposed immunosensor to AFB1 using homology modeling combined with molecular docking. In the presence of AFB1, the current signal of the modified electrode reduced; this involved specific antibody-antigen binding, including hydrophobic hydrogen bonding and pi-pi stack interactions. The new AFB1 sensor platform showed two linearity ranges of 0.01-1 ng mL-1 and 1-100 ng mL-1, with the limit of detection at 0.0019 ng mL-1. We investigated the proposed immunosensor in real samples, including peanuts, certified reference material of a peanut sample (labeled 206 µg kg-1 AFB1), corn, and chicken feed. The sensor's accuracy was 86.1-104.4% recovery, which agrees with the reference HPLC technique using paired t-test analysis. The present work shows excellent performance for AFB1 detection and could be applied for food quality control or modified to detect other mycotoxins.

Ultrasensitive electrochemiluminescence sensor based on nitrogen-decorated carbon dots for Listeria monocytogenes determination using a screen-printed carbon electrode.

Current method for identification of foodborne pathogens suffers from its relatively poor performance, consequently limiting its use. Herein, we first describe an ultrasensitive electrochemiluminescence (ECL) sensor based on nitrogen-decorated carbon dots (NCDs) for Listeria monocytogenes (L. monocytogenes) determination using a screen-printed carbon electrode (SPCE). Citric acid serves as carbon source, and ethylenediamine, a molecule containing nitrogen atom, is employed to synthesize CDs. Approximately 4 nm NCD with homogenous size distribution can be produced via a single step green microwave-assisted methodology. The construction of ECL sensor is initiated by the immobilization of capture antibody (Ab1) onto the carboxyl graphene (GOOH)-modified SPCE, where immunocomplexes (antigen and the NCD-labelled secondary antibody (Ab2-NCD)) are formed, resulting in a substantial increment in the ECL signal response in the presence of K2S2O8. The GOOH allows direct formation of the capture antibodies and enhances the electrochemical properties. Under optimal parameters, this sensor exhibits wide linearity (2 to 1.0 × 106 CFU mL-1), high sensitivity (0.104 or 1.0 × 10-1 CFU mL-1) and specificity over the nontargeting studied pathogens and is successfully applied to determine L. monocytogenes in food products. These promising results together with its performance suggest that this proposed platform may serve as an alternative device to effectively control the spread of foodborne diseases.

A portable selective electrochemical sensor amplified with Fe3O4@Au-cysteamine-thymine acetic acid as conductive mediator for determination of mercuric ion.

Mercury ion (Hg2+) is considered to be one of the most toxic heavy metal ions and can cause adverse effects on kidney function, the central nervous system, and the immune system. Therefore, it is important to develop a fast and simple method for sensitive and selective detection of Hg2+ in the environment. This research proposes a portable electrochemical sensor for rapid and selective detection of Hg2+. The sensor platform is designed based on thymine acetic acid anchored with cysteamine-conjugated core shell Fe3O4@Au nanoparticles (Fe3O4@Au/CA/T-COOH) immobilized on a sensing area of a screen-printed carbon electrode (SPCE) with the aid of an external magnetic field embedded in a homemade electrode holder for ease of handling. In the presence of Hg2+, the immobilized thymine combines specifically with Hg2+ and forms a thymine-Hg2+-thymine mismatch (T-Hg2+-T). The resulting amount of Hg2+ was determined by differential pulse anodic stripping voltammetry (DPASV). Under optimal conditions, the sensor exhibited two wide linearities in a range from 1 to 200 µg L-1 and 200-2200 µg L-1 with the reliability coefficient of determination of 0.997 and 0.999, respectively. The detection limit (LOD) and the quantification limit (LOQ) were also determined to be 0.5 µg L-1 and 1.0 µg L-1, respectively. The sensor was further applied for determination of Hg2+ in water samples, a certified reference material and fish samples. The results were compared with flow injection atomic spectroscopy-inductively coupled plasma-optical emission spectroscopy (FIAS-ICP-OES) systems as a reference method. Results obtained with the proposed sensor were relatively satisfactory, and they showed no significant differences at a 95% confidence level by t-test from the standard method. Therefore, considering its fast and simple advantages, this novel strategy provides a potential platform for construction of a Hg2+ electrochemical sensor.

Tungsten - Tungsten Trioxide Electrodes for the Long-term Monitoring of Corrosion Processes in Highly Alkaline Media and Concrete-based Materials.

The determination of pH in highly alkaline solutions and concrete materials is extremely important for monitoring or predicting the corrosion processes of reinforced concrete structures and to follow the hydration process of Portland cement, fly-ash, micro silica and other materials used in concrete manufacturing. The corrosion of reinforced concrete structures and the hydration of pozzolanic materials are long-term processes, which means, that appropriate durable, and resilient pH electrodes are needed, for direct implantation regarding solid concrete bodies. The purpose of this work was to characterise the potentiometric and surface properties of tungsten electrodes after exposure to extreme alkaline solutions. The tungsten wire surface was activated at 800 °C for 30 min within an oxygen flow. The formation of homogenous and compact multiple layers of WO3 crystals was observed using X-ray diffraction and scanning electron microscopy. X-ray diffraction of those tungsten electrodes exposed to saturated calcium hydroxide solution or the pore-water of cement-based materials during 10 months, indicated partly dissolved WO3. Two new compounds appeared on the electrodes surfaces; pure tungsten and CaWO4. The presence of tungsten was affecting any potentiometric response in acidic pH region (2-5) but in pH 5-12 region the response still remained linear with a slope of 42 ± 2 mV/pH unit. The W/WO3 electrode was suitable for the long-term monitoring of corrosion processes in concrete-based materials according to the pH changes as it has stable and repeatable responses to alkaline solutions (pH > 12). All the tested interferring ions had no significant influence on electrode potential. The W/WO3 electrode is simple, robust, inexpensive, and temperature resistant and can be applied in potentiometric titrations as well as in batch and flow-injection analysis. The prepared electrode is a highly promising pH sensor for the monitoring of pH changes in highly alkaline capillary water of concrete.

An ultrasensitive immunosensor based on manganese dioxide-graphene nanoplatelets and core shell Fe3O4@Au nanoparticles for label-free detection of carcinoembryonic antigen.

A novel electrochemical immunosensor was developed for label-free detection of carcinoembryonic antigen (CEA) as a cancer biomarker. The designed immunosensor was based on CEA antibody (anti-CEA) anchored with core shell Fe3O4@Au nanoparticles which were immobilized on a screen-printed carbon electrode modified with manganese dioxide decorating on graphene nanoplatelets (SPCE/GNP-MnO2/Fe3O4@Au-antiCEA). The SPCE was placed onto a home-made electrode holder for easy handling. The approach was based on direct binding of CEA to a fixed amount of anti-CEA on the modified electrode for the specific detection using linear sweep voltammetry (LSV) and electrochemical impedance spectroscopy (EIS) monitored in a solution containing 5 mM [Fe(CN)63-/4-] prepared in 0.1 M phosphate buffer at pH 7.4. The difference in signal response owing to the redox reaction of [Fe(CN)6]3-/4- before and after interaction with CEA was regarded as the immunosensor response corresponding directly to the CEA concentration. Under optimized conditions, the linear range of 0.001-100 ng/mL, and the detection limits of 0.10 pg/mL (LSV) and 0.30 pg/mL (EIS) were evaluated. The applicability of the immunosensor was verified by well-corresponding determination of CEA in diluted human serum samples by electrochemiluminescence (ECL) immunoassay. Therefore, the proposed immunosensor could be suitable enough for a real sample analysis of CEA.

Electrochemical immunosensor functionalized with nanobodies for the detection of the toxic microalgae Alexandrium minutum using glassy carbon electrode modified with gold nanoparticles.

In this work an electrochemical immunosensor for the toxic microalgae Alexandrium minutum (A. minutum AL9T) detection is described. A glassy carbon electrode (GCE) was modified by depositing gold nanoparticles followed by L-cysteine for obtaining a self-assembled monolayer. The SpyTagged nanobody C1, specific for the A. minutum toxic strain AL9T, was then covalently immobilized via SpyCatcher on the surface of the modified electrode and used for the selective capture of such microalgae strain. Electrochemical impedance spectroscopy (EIS) was used for the quantification of A. minutum cells present in water samples by measuring the charge-transfer resistance changes of the electrode with a hexacyanoferrate probe. Each electrode modification step was accompanied by cyclic voltammetry (CV) and scanning electron microscopy (SEM). The immunosensor provided highly reproducible data, was simple to fabricate at low cost, exhibited higher sensitivity than previously described alternative diagnostic methods and showed a broad linear range between 103 and 109 cells L-1 with detection limit of 3 × 103 cells L-1 of A. minutum AL9T. The immunosensor was successfully applied to quantify A. minutum AL9T in seawater and brackish water samples proving that it can be used for early detection of harmful microalgae without the necessity of pre-concentration or dialysis steps.