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An ultrasensitive impedimetric glycan-based biosensor for reliable and selective detection of inactivated, but intact influenza viruses H3N2 was developed. Such glycan-based approach has a distinct advantage over antibody-based detection of influenza viruses since glycans are natural viral receptors with a possibility to selectively distinguish between potentially pathogenic influenza subtypes by the glycan-based biosensors. Build-up of the biosensor was carefully optimized with atomic force microscopy applied for visualization of the biosensor surface after binding of viruses with the topology of an individual viral particle H3N2 analyzed. The glycan biosensor could detect a glycan binding lectin with a limit of detection (LOD) of 5 aM. The biosensor was finally applied for analysis of influenza viruses H3N2 with LOD of 13 viral particles in 1 μl, what is the lowest LOD for analysis of influenza viral particles by the glycan-based device achieved so far. The biosensor could detect H3N2 viruses selectively with a sensitivity ratio of 30 over influenza viruses H7N7. The impedimetric biosensor presented here is the most sensitive glycan-based device for detection of influenza viruses and among the most sensitive antibody or aptamer based biosensor devices. [ABSTRACT FROM AUTHOR]

A novel fabrication process for composite consisting of organized modified carbon nanotubes (CNT) in the polyacrylic acid (PAA) matrix has been developed. The CNT were organized along crystallite domains during a directional freezing process and subsequently fixed by the gamma irradiation induced polymerization and crosslinking in solid state. Different morphology and distribution of CNT in fabricated and conventionally prepared composites were investigated by scanning and transmission electron microscopies. The interactions in the polymer matrix were elucidated by a broad dielectric spectroscopy. Importantly, the composite exhibited anisotropic electrical and mechanical properties studied by electrical conductivity measurement and dynamic mechanical analysis, respectively. Differences of two orders of magnitude in the electrical conductivity and 5 times higher storage modulus at 10 − 2 Hz in the direction along and perpendicular to freezing in composite with 0.1% CNT in feed were observed. [ABSTRACT FROM AUTHOR]

In this study, an assay for detection of the cancer biomarker Thomsen-nouvelle (Tn) antigen on the ELISA plates format was designed and developed. The effects of size and the interfacial density of the negative charge of magnetic beads (MBs) on the specific sensitivity of the bioaffinity interaction were studied. In particular, glyconanoconjugate, i.e. glycan Tn antigen conjugated to bovine serum albumin (BSA) was covalently immobilised on MBs for the bioaffinity detection of anti-Tn antibodies as cancer biomarkers. Six different MBs were used in the study, i.e. carboxy-modified MBs of 250 nm, 500 nm, 1000 nm and 2800 nm and epoxy-modified MBs of 2800 nm and 4500 nm. In order to evaluate which MBs are the best suited for detection of the analyte anti-Tn antibodies, sensitivities of detection (slopes from calibration curves) were calculated. Next, specific sensitivities were calculated for each type of MBs as a ratio of sensitivity of detection to the mass of MBs. From zeta potential ζ for each type of MBs, the interfacial charge density on MBs was calculated, expressed as the density of zeta potential ζ d (ratio of zeta potential to surface area of MBs, i.e. ζ d = ζ/A). Then, we evaluated the effect of size and ζ d on the specific sensitivity of detection of anti-Tn antibodies in order to understand the immobilisation process on nanoscale. We also identified an optimal value of ζ d on MBs; this was essential to achieve highly sensitive detection of the analyte, which made it possible to attain limit of detection (LOD) of (0.31 ± 0.01) ng mL−1 or (2.10 ± 0.04) pM for analyte detection. In addition, the optimal assay configuration was highly selective and enabled reliable detection of the analyte in human serum with a recovery index in the range of 102–104%. Diffusion of negatively charged glyconanoconjugate to magnetic beads with different density of negative charge [Display omitted] • Immobilisation of a glyconanonjugate on magnetic beads (MBs) with different size and functionalities. • Detection of anti-glycan antibodies (AGA) using MBs with glyconanoconjugate in ELISA format. • Calculation of interfacial charge density on MBs as the density of zeta potential ζ d. • The highest specific sensitivity for detection of AGA was achieved on MBs with low ζ d. • AGA was detected with LOD of (0.31 ± 0.01) ng mL−1 or (2.10 ± 0.04) pM with recovery index of 102-104%. [ABSTRACT FROM AUTHOR]

In this study, we applied MXene as column cartridge for N -glycan enrichment from human samples with a focus on the analysis of sialic acid linkages using a derivatisation protocol followed by glycan analysis via Matrix Assisted Laser Desorption Ionisation-Time Of Flight Mass Spectrometry (MALDI-TOF-MS). The MXene-based cartridge enriches a higher number of glycans (i.e., sialylated and bisecting N -glycans) when compared to the commercial HILIC columns. We demonstrate the strong potential of MXene as a stationary phase in MS glycomic analysis. [Display omitted] • N -glycan enrichment by a traditional Ludger column from human serum sample. • N -glycan enrichment by a novel 2D nanomaterial MXene based column from human serum sample. • Comparison of both enrichment strategies shows better performance of MXene-base enrichment. • 2D nanomaterial MXene is able to enrich sialic acid containing glycans and bisecting glycans, which are cancer associated. [ABSTRACT FROM AUTHOR]

Systemic sclerosis (SSc) is an autoimmune disease seriously affecting patient’s quality of life. The heterogeneity of the disease also means that identification and subsequent validation of biomarkers of the disease is quite challenging. A fully validated single biomarker for diagnosis, prognosis, disease activity and assessment of response to therapy is not yet available. The main aim of this study was to apply an alternative assay protocol to the immunoassay-based analysis of this disease by employment of sialic acid recognizing lectin Sambucus nigra agglutinin (SNA) to glycoprofile serum samples. To our best knowledge this is the first study describing direct lectin-based glycoprofiling of serum SSc samples. Three different analytical methods for glycoprofiling of serum samples relying on application of lectins are compared here from a bioanalytical point of view including traditional ELISA-like lectin-based method (ELLA), novel fluorescent lectin microarrays and ultrasensitive impedimetric lectin biosensors. Results obtained by all three bioanalytical methods consistently showed differences in the level of sialic acid present on glycoproteins, when serum from healthy people was compared to the one from patients having SSc. Thus, analysis of sialic acid content in human serum could be of a diagnostic value for future detection of SSc, but further work is needed to enhance selectivity of assays for example by glycoprofiling of a fraction of human serum enriched in antibodies for individual diagnostics. [ABSTRACT FROM AUTHOR]

In this article we describe construction of a bioreceptive interface for detection of a breast cancer biomarker carbohydrate antigen CA15-3. The conductive interface was patterned by a 2D nanomaterial MXene, to which a mixed layer containing sulfobetaine and carboxybetaine was electrochemically grafted through a diazonium moiety. Such a modified interface was then applied for covalent immobilisation of anti-CA15-3 antibody as a bioreceptive probe for detection of a breast cancer biomarker. Two different strategies were applied for final construction of an immunosensor i.e. an interface finally blocked by bovine serum albumin or an immunosensor without such modification. Finally, electrochemical reading was accomplished using a soluble redox probe Ru(NH 3) 6 3+ ion for detection of CA15-3 in a clinically relevant range up to 50 U mL−1. The results indicate that immunosensor based on non-blocked interface can be applied for biosensing using two modes of action: 1. differential pulse voltammetry (a plot of a peak current vs. analyte concentration) and 2. an electrochemical impedance spectroscopy (a plot of a charge transfer resistance vs. analyte concentration). The electrode blocked by bovine serum albumin (BSA) can be used by additional 3. mode of action: through detection of changes in the potential (a plot E p vs. c). Additionally, we reveal and explain that Ru(NH 3) 6 3+ is redox probe, which can be applied as interfacial molecular nanoscale ruler to distinguish negatively charged protein molecules present in the close proximity (≤ 6 nm) of the electrode (in our case adsorbed BSA molecules) from the negatively charged protein molecules at a larger distance (>12 nm) from the electrode (i.e. CA15-3 analyte). Two ways of bioaffinity interactions using on a conductive surface non-blocked by bovine serum albumin (BSA) (A) and blocked by BSA (B). [Display omitted] • Modification of glassy carbon electrode by the 2D nanomaterial MXene. • Patterning of GCE/MXene interface by a mixed layer and zwitterions by electrochemical trigger (non-blocked electrodes). • Further modification of the electrode by bovine serum albumin (blocked electrodes). • Examination of differences in the electrochemical behaviour of blocked and non-blocked electrodes. • Three modes of biosensing by electrochemical means possible using EIS, DPV and CV assays using Ru(NH 3) 6 3+ ion redox probe. [ABSTRACT FROM AUTHOR]

[Display omitted] • Electrochemical activation by ORR of commercial tungsten carbide was done. • Induction of oxycarbides species on surface activated WC material. • In-situ study identified improvement inchemisorption. • Enhanced electrocatalytic performance achieved by processing activation. The chemistry of electrocatalysts deals with multiple critical factors to facilitate the electrochemical reactions. Among those, the rate limiting depends on electrons transfer for chemisorptions of molecules in redox reactions. This feature can be directly linked with efficient catalyst support material in electrocatalysis. To this end, we have developed a novel, simple and facile route to introduce commercially available material with tuned surface and interface chemistry for their potential applications in fuel cells (FCs) science and technology. Commercial tungsten carbide (WC) was activated by means of electrochemical oxygen reduction reactions (ORR) on different rotation rates to induce mild interactions of oxygen molecules with surface of WC at specified reduction potentials. The X-ray diffraction and X-ray photoelectron spectroscopy analysis before and after the activation confirmed the tuning of WC surface with incorporation of potential factors to activate them for enhanced electrocatalytic activities. In addition, the electrocatalytic methanol oxidation reactions (MOR) and hydrogen evolution reactions (HER) were carried and confirmed the exceptional boosted-up electrocatalytic behaviour of WC after the activation. The enhancement in electrocatalytic mechanism after activation was also tested and proved by means of in-situ FTIR spectroelectrochemical analysis for methanol electro-oxidation. In addition, the electrochemical depositions of Pt nanoparticles were carried out on WC surface before and after the activation to reveal the influence of surface activation for accommodating the foreign particles as support material in electrocatalysis. The results shown two fold enhancement in anodic performance of Pt-modified activated WC catalyst for methanol oxidation reactions and hydrogen evolution reactions in fuel cells. [ABSTRACT FROM AUTHOR]

Exosomes are considered to be a rich source of biomarkers, hence in this article we examine the best procedure for their isolation. We examine several isolation procedures, exosome storage conditions and other conditions affecting exosome production by prostate cell lines. We selected four different commercially available kits based on different principles to achieve exosome isolation, the best being magnetic-based. In addition, we found storage at − 20 °C to be good for storing isolated exosomes and that exosomes were produced from the cancerous prostate cell line 22Rv1 in much greater amounts than the non-cancerous prostate cell line RWPE1. We also found differences in the response of both cell lines in the production of exosomes as a result of stress, i.e. exposure to hydrogen peroxide and starvation. The effect of Triton X-100 on exosome lysis was examined using two different surfactant concentrations by analysis of the exosome count and change in the exosome size. The final part of the article details the advantages of the use of a 2D biochip prepared in-house over a commercially available 3D biochip for monitoring the interaction of exosomes via its surface receptors (CD63) with an immobilised ligand (anti-CD63 antibodies) using surface plasmon resonance. The final experiment shows the potential of lectin fluorescent microarrays for the analysis of glycans present in lysed exosomes. [Display omitted] • Comparison of four exosome isolation kits. • Examination of storage conditions on exosomes. • Comparison of exosome production rate between prostate control and prostate cancerous cell line. • Evaluation of starvation and oxidative stress on cells ability to produce exosomes. • Comparison of exosome analysis by SPR using 2D and 3D interfaces [ABSTRACT FROM AUTHOR]

We present here an ultrasensitive electrochemical biosensor based on a lectin biorecognition capable to detect concentrations of glycoproteins down to attomolar (aM) level by investigation of changes in the charge transfer resistance (R ct ) using electrochemical impedance spectroscopy (EIS). On polycrystalline gold modified by an aminoalkanethiol linker layer, gold nanoparticles were attached. A Sambucus nigra agglutinin was covalently immobilised on a mixed self-assembled monolayer formed on gold nanoparticles and finally, the biosensor surface was blocked by poly(vinyl alcohol). The lectin biosensor was applied for detection of sialic acid containing glycoproteins fetuin and asialofetuin. Building of a biosensing interface was carefully characterised by a broad range of techniques such as electrochemistry, EIS, atomic force microscopy, scanning electron microscopy and surface plasmon resonance with the best performance of the biosensor achieved by application of HS–(CH2)11–NH2 linker and gold nanoparticles with a diameter of 20nm. The lectin biosensor responded to an addition of fetuin (8.7% of sialic acid) with sensitivity of (338±11) Ωdecade−1 and to asialofetuin (≤0.5% of sialic acid) with sensitivity of (109±10) Ωdecade−1 with a blank experiment with oxidised asialofetuin (without recognisable sialic acid) revealing sensitivity of detection of (79±13) Ωdecade−1. These results suggest the lectin biosensor responded to changes in the glycan amount in a quantitative way with a successful validation by a lectin microarray. Such a biosensor device has a great potential to be employed in early biomedical diagnostics of diseases such as arthritis or cancer, which are connected to aberrant glycosylation of protein biomarkers in biological fluids. [ABSTRACT FROM AUTHOR]

Abstract: Spherical carbon nanoparticles KetjenBlack (KB) with a high sorption capacity together with conductivity increasing single walled carbon nanotubes (CNTs) were “glued” together by chitosan for the preparation of a composite. A biocathode with bilirubin oxidase (BOD) adsorbed within the composite was characterised and its composition optimised. A renewable biopolymer chitosan present in the composite offered (1) pre-concentration of BOD within the matrix via electrostatic interactions, (2) favourable orientation of BOD for a direct electron transfer (DET) between BOD and the composite, (3) electrochemical visibility of 3 redox sites present in BOD, (4) low charge transfer resistance, (5) high proton conductivity and (6) low overpotential for oxygen reduction. Electrochemical investigation of BOD revealed interesting internal redox communication within the enzyme with some novel insights provided. At least one of tyrosines present in BOD seems to be involved in electron transfer route, as well. The composite used for the biocathode was directly applied for the preparation of a bioanode with fructose dehydrogenase immobilised, working in a DET mode of operation. Finally, integration of the biocathode and the bioanode into a biofuel cell operated in a reagentless and membraneless mode offered a power density of 50μWcm−2 at 300mV. [Copyright &y& Elsevier]

We report on an ultrasensitive label-free lectin-based impedimetric biosensor for the determination of the sialylated glycoproteins fetuin and asialofetuin. A sialic acid binding agglutinin from Sambucus nigra I was covalently immobilised on a mixed self-assembled monolayer (SAM) consisting of 11-mercaptoundecanoic acid and 6-mercaptohexanol. Poly(vinyl alcohol) was used as a blocking agent. The sensor layer was characterised by atomic force microscopy, electrochemical impedance spectroscopy and X-ray photoelectron spectroscopy. The biosensor exhibits a linear range that spans 7 orders of magnitude for both glycoproteins, with a detection limit as low as 0.33 fM for fetuin and 0.54 fM for asialofetuin. We also show, by making control experiments with oxidised asialofetuin, that the biosensor is capable of quantitatively detecting changes in the fraction of sialic acid on glycoproteins. We conclude that this work lays a solid foundation for future applications of such a biosensor in terms of the diagnosis of diseases such as chronic inflammatory rheumatoid arthritis, genetic disorders and cancer, all of which are associated with aberrant glycosylation of protein biomarkers. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Glycans and other saccharide moieties attached to proteins and lipids, or present on the surface of a cell, are actively involved in numerous physiological or pathological processes. Their structural flexibility (that is based on the formation of various kinds of linkages between saccharides) is making glycans superb "identity cards". In fact, glycans can form more "words" or "codes" (i.e., unique sequences) from the same number of "letters" (building blocks) than DNA or proteins. Glycans are physicochemically similar and it is not a trivial task to identify their sequence, or-even more challenging-to link a given glycan to a particular physiological or pathological process. Lectins can recognise differences in glycan compositions even in their bound state and therefore are most useful tools in the task to decipher the "glycocode". Thus, lectin-based biosensors working in a label-free mode can effectively complement the current weaponry of analytical tools in glycomics.This review gives an introduction into the area of glycomics and then focuses on the design, analytical performance, and practical utility of lectin-based electrochemical label-free biosensors for the detection of isolated glycoproteins or intact cells. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Abstract: A biocompatible nanocomposite consisting of single-walled carbon nanotubes (CNTs) dispersed in a hyaluronic acid (HA) was investigated as a sensing platform for a mediatorless electrochemical detection of NADH. The device was characterised by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and extensively by electrochemistry. CNT–HA bionanocomposite showed more reversible electrochemistry, higher short-term stability of NADH sensing and higher selectivity of NADH detection compared to frequently used CNT–CHI (chitosan) modified GCE. Finally the performance of the sensor modified by CNT–HA was tested in a batch and flow injection analysis (FIA) mode of operation with basic characteristics revealed. The NADH sensor exhibits a good long-term operational stability (95% of the original sensitivity after 22h of continuous operation). Subsequently a d-sorbitol biosensor based on such a nanoscale built interface was prepared and characterised with a d-sorbitol dehydrogenase used as a biocatalyst. [Copyright &y& Elsevier]

Abstract: A ferricyanide mediated amperometric biosensor system implementing d-sorbitol dehydrogenase together with diaphorase for sensitive detection of d-sorbitol was used. The biosensor system was successfully integrated into an off-line FIA system with a throughput of detection of 10h−1. The device exhibited limit of detection of 20μM with an average relative standard deviation of analysis of samples of 2.2%. The signal of the biosensor was linear up to 1.1mM for d-sorbitol with sensitivity of (72±2)nAmM−1, while a dynamic range was much wider up to 18mM. The sorbitol biosensor gave reliable results even in the presence of a high molar excess of l-sorbose, a product of the biotransformation process, as judged from an excellent agreement with HPLC and GC. [Copyright &y& Elsevier]

The development of a novel SUspension Magnetic-Bead-based Assay (SUMBA) for the detection of antibodies against aberrant glycans (AGA) as potential cancer biomarkers is presented here. The SUMBA method was extensively optimised by choosing proper commercially available AGA able to specifically, and with high affinity, recognise aberrant glycans, which were attached to the protein backbone working as a molecular scaffold (a glycoconjugate). The whole SUMBA was optimised using several analytical techniques such as Surface Plasmon Resonance and Energy Dispersive X-ray Spectroscopy. Additionally, the SUMBA method was extensively optimised for signal enhancement. With all steps optimised, we were able to detect AGA ultrasensitively with a limit of detection of 0.45 pM. Moreover, AGA could be detected in serum samples with a recovery index in the range of 98%–104%. SUMBA applied for detection of anti-glycan antibodies in serum samples. [Display omitted] • Application of magnetic beads modified with aberrant glycans for enrichment of AGA. • Use of polyHRP for signal enhancement. • A direct comparison of magnetic bead-based assay to ELISA method of detection. • LOD of 450 fM and recovery index of 98–104% for detection of anti-Tn AGA in serum sample. [ABSTRACT FROM AUTHOR]

Principal component analysis applied for clustering of patients with benign prostatic hyperplasia (left) and patients with prostate cancer (right). Outside the cluster are patients with changed glycosylation of free form of prostate specific antigen compared to the rest of the patients in the cohort. PCA applied for identification of castration resistant prostate cancer patients in the BPH cohort. [Display omitted] • Analysis of changes in the glycan composition of free form of prostate specific antigen (fPSA) was done in ELISA format. • Individual and Double biomarkers were applied to discriminate benign prostate hyperplasia patients (BPH) from prostate cancer (PCa) patients. • The best double biomarker is gPSA (glycan form of fPSA) and PSAd (PSA/prostate volume) • Such biomarker offers high discrimination power BPH vs. PCa with AUC (area under receiver operating curve) of 0.896. • Genetic algorithm was applied to identify false negatives in the benign cohort. • Principal component analysis (PCA) applied for identification of prostatis patient in the BPH cohort; Prostate cancer (PCa) is one of the most common cancer types among men and also a common cause of death globally. With an increasing incidence, there is a need for low-cost, reliable biomarkers present in samples, which could be provided non-invasively (without a need to perform prostate biopsy). Glycosylation changes of free-PSA (fPSA) are considered cancer-specific, while the level of different PSA forms can increase under other than cancerous conditions. In the present study, we investigated the role of N , N -diacetyllactosamine (LacdiNAc) epitope of fPSA (i.e. glycoprofile of fPSA or gPSA) in combination with total-PSA (tPSA), prostate volume, and tPSA density (tPSA level divided by prostate volume i.e. PSAd) as biomarkers for monitoring of PCa development and progression in 105 men. Furthermore, we applied an genetic (evolutionary) algorithm to identify any suspicious individuals in a benign cohort having benign prostatic hyperplasia (BPH). We identified 3 suspicious men originally diagnosed with BPH using gPSA analysis. In the follow-up we found out that two men should not be considered as BPH patients since multiparametric magnetic resonance imaging (mpMRI) identified one man with clinically significant PCa via Prostate Imaging – Reporting and Data System (PI RADS v2 = 4) and the second man was with High-grade prostatic intraepithelial neoplasia (HG PIN), commonly described as a pre-cancerous stage. Moreover, in the study we described for the first time that changed LacdiNAc on PSA can be applied to identify prostatitis patients and most importantly this is the first study suggesting that changed glycosylation on PSA can be applied to identify castration-resistant prostate cancer (CRPCa) patients. [ABSTRACT FROM AUTHOR]

The review article provides a short introduction to exosomes with the focus to use exosomes as disease markers itself (i.e. their concentration or presence of some specific receptors) or a source of disease biomarkers such as proteins and metabolites. In detail, we are discussing various methods of exosome isolation and the main focus of the review paper is on affinity capture of exosomes, since some of them can be applied to the isolation of specific sub-populations of exosomes produced by some specific organs. The article provides a comprehensive overview of magnetic (bio)affinity capture applied to the detection of exosomes or exosomal cargo using different (bio)affinity capture ligands such as antibodies, DNA aptamers, peptides, glycan-based recognition, transferrin-based approaches, affinity based on recognition of phospholipids of exosomes and other approaches including electrostatic interactions. The review in detail provides key analytical and clinical parameters of such approaches in a form of an extensive table summarising outcomes published in the last two years (2023-2024). Finally, the review paper also provides conclusions sections discussing pros and cons of magnetic (bio)affinity capture for exosome isolation and/or determination  of exosomal content., Competing Interests: Declarations. Conflict interests: The authors declare no competing interests., (© 2025. The Author(s).)

Post-translational modifications of proteins play an important role in their stability, solubility and in vivo function. Also, for several reasons, such as the Golgi fragmentation during cancerogenesis, glycosylation as the most common modification is especially promising in offering high cancer specificity which, in combination with tissue-specific biomarkers available in the case of prostate diseases (PSA, PSMA, PAP), may lead to the development of novel oncodiagnostic approaches. In this review, we present the importance of subterminal glycan structures based on the N-acetylated monosaccharides GlcNAc and GalNAc in N- and also O-glycans, structures of which they are a component (LacNAc, LacdiNAc, branched structures). We also discuss the importance and clinical performance of these structures in cases of prostate cancer diagnostics using lectin-based affinity methods, which could be implemented in clinical laboratory practice in the future.

Screen-printing technology is a game changer in many fields including electrochemical biosensing. Two-dimensional nanomaterial MXene Ti 3 C 2 T x was integrated as a nanoplatform to immobilise enzyme sarcosine oxidase (SOx) onto the interface of screen-printed carbon electrodes (SPCEs). A miniaturised, portable, and cost-effective nanobiosensor was constructed using chitosan as a biocompatible glue for the ultrasensitive detection of prostate cancer biomarker sarcosine. The fabricated device was characterised with energy-dispersive X-ray spectroscopy (EDX), electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). Sarcosine was detected indirectly via the amperometric detection of H 2 O 2 formed during enzymatic reaction. The nanobiosensor could detect sarcosine down to 7.0 nM with a maximal peak current output at 4.10 ± 0.35 × 10 -5 A using only 100 µL of a sample per measurement. The assay run in 100 μL of an electrolyte showed the first linear calibration curve in a concentration window of up to 5 μM with a slope of 2.86 μA·μM -1 , and the second linear calibration curve in the range of 5-50 μM with a slope of 0.32 ± 0.01 μA·μM -1 (R 2 = 0.992). The device provided a high recovery index of 92.5% when measuring an analyte spiked into artificial urine, and could be used for detection of sarcosine in urine for at least a period of 5 weeks after the preparation.

Colorectal cancer (CRC) is one of the most common types of cancer among men and women worldwide. Efforts are currently underway to find novel and more cancer-specific biomarkers that could be detected in a non-invasive way. The analysis of aberrant glycosylation of serum glycoproteins is a way to discover novel diagnostic and prognostic CRC biomarkers. The present study investigated a whole-serum glycome with a panel of 16 different lectins in search for age-independent and CRC-specific glycomarkers using receiver operating characteristic (ROC) curve analyses and glycan heat matrices. Glycosylation changes present in the whole serum were identified, which could lead to the discovery of novel biomarkers for CRC diagnostics. In particular, the change in the bisecting glycans (recognized by Phaseolus vulgaris erythroagglutinin) had the highest discrimination potential for CRC diagnostics in combination with human L selectin providing area under the ROC curve (AUC) of 0.989 (95% CI 0.950-1.000), specificity of 1.000, sensitivity of 0.900, and accuracy of 0.960. We also implemented novel tools for identification of lectins with strong discrimination power., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Bertok, Bertokova, Jane, Hires, Aguedo, Potocarova, Lukac, Vikartovska, Kasak, Borsig and Tkac.)

In this study, we propose a new approach to attain energy by salinity gradient engines with pistons based on hydrogels possessing polyelectrolyte and antipolyelectrolyte effects in a tandem arrangement, providing energy in each salinity gradient mode in a repeatable manner. The swelling of hydrogel with a polyelectrolyte effect and shrinking of hydrogel particles possessing an antipolyelectrolyte effect in desalinated water, and subsequent shrinking of hydrogel with polyelectrolyte and swelling of hydrogel antipolyelectrolyte effect in saline water, generate power in both increasing and decreasing salinity modes. To investigate the energy recovery, we scrutinized osmotic engine assemblies by a setup arrangement of pistons with hydrogel particles, with polyelectrolyte and antipolyelectrolyte effects, in tandem. The energy recovery from the tandem engine setup (calculated based on dry form for each polyelectrolyte polyacrylate-based hydrogel-SPA) and antipolyelectrolyte-sulfobetaine-based gel with methacrylate polymeric backbone-SBE) up to 581 J kg -1 and a mean power of 0.16 W kg -1 was obtained by the tandem setup of SPA and SBE hydrogel containing 3% crosslinking density and particle size of 500 microns with an external load of 3.0 kPa. Exchange of sulfobetaine with methacrylamide (SBAm), the main polymer backbone, revealed a positive increase in energy recovery of 670 J kg -1 with a mean power of 0.19 W kg -1 for the tandem system operating under the same parameters (SPA@SBAm). The energy recovery can be controlled, modulated and tuned by selecting both hydrogels with antipolyelectrolyte and polyelectrolyte effects and their performing parameters. This proof of concept provides blue energy harvesting by contributing both polyelectrolyte and antipolyelectrolyte effects in a single tandem setup; together with easy accessibility (diaper-based materials (SPA)) and known antibiofouling, these properties offer a robust alternative for energy harvesting.

Introduction: Breast cancer (BCa) is the most common cancer type diagnosed in women and 5 th most common cause of deaths among all cancer deaths despite the fact that screening program is at place. This is why novel diagnostics approaches are needed in order to decrease number of BCa cases and disease mortality., Areas Covered: In this review paper, we aim to cover some basic aspects regarding cellular metabolism and signalling in BCa behind altered glycosylation. We also discuss novel exciting discoveries regarding glycan-based analysis, which can provide useful information for better understanding of the disease. The final part deals with clinical usefulness of glycan-based biomarkers and the clinical performance of such biomarkers is compared to already approved BCa biomarkers and diagnostic tools based on imaging., Expert Opinion: Recent discoveries suggest that glycan-based biomarkers offer high accuracy for possible BCa diagnostics in blood, but also for better monitoring and management of BCa patients. The review article was written using Web of Science search engine to include articles published between 2019 and 2021.

This is the first work focused on glycoprofiling of whole N- and O- glycome using lectins in an array format applied for analysis of serum samples from healthy individuals, benign prostate hyperplasia (BPH) patients, and prostate cancer (PCa) patients. Lectin microarray was prepared using traditional lectins with the incorporation of 2 recombinant bacterial lectins and 3 human lectins (17 lectins in total). Clinical validation of glycans as biomarkers was done in two studies: discrimination of healthy individuals with BPH patients vs. PCa patients (C vs. PCa) and discrimination of healthy individuals vs. BPH and PCa patients (H vs. PCond). Single lectins (17 lectins) and a combination of two lectins (136 binary lectin combinations) were applied in the clinical validation of glycan biomarkers providing 153 AUC values from ROC curves for both studies (C vs. PCa and H vs. PCond). Potential N- and O-glycans as biomarkers were identified and possible carriers of these glycans are shortly discussed.

For efficient and effective utilization of MXene such as biosensing or advanced applications, interfacial modification of MXene needs to be considered. To this end, we describe modification of Ti 3 C 2 T x MXene by aryldiazonium-based grafting with derivatives bearing a sulfo- (SB) or carboxy- (CB) betaine pendant moiety. Since MXene contains free electrons, betaine derivatives could be grafted to MXene spontaneously. Kinetics of spontaneous grafting of SB and CB toward MXene was electrochemically examined in two different ways, and such experiments confirmed much quicker spontaneous SB grafting compared to spontaneous CB grafting. Moreover, a wide range of electrochemical methods investigating non-Faradaic and Faradaic redox behavior also in the presence of two redox probes together with contact-angle measurements and secondary ion mass spectrometry (SIMS) confirmed substantial differences in formation and interfacial presentation of betaine layers, when spontaneously grafted on MXene. Besides spontaneous grafting of CB and SB toward MXene, also electrochemical grafting by a redox trigger was performed. Results suggest that electrochemical grafting provides a denser layer of SB and CB on the MXene interface compared to spontaneous grafting of SB and CB. Moreover, an electrochemically grafted SB layer offers much lower interfacial resistance and an electrochemically active surface area compared to an electrochemically grafted CB layer. Thus, by adjusting the SB/CB ratio in the solution during electrochemical grafting, it is possible to effectively tune the redox behavior of an MXene-modified interface. Finally, electrochemically grafted CB and SB layers on MXene were evaluated against non-specific protein binding and compared to the anti-fouling behavior of an unmodified MXene interface., (Copyright © 2020 Lorencova, Gajdosova, Hroncekova, Bertok, Jerigova, Velic, Sobolciak, Krupa, Kasak and Tkac.)

Two-dimensional layered nanomaterial Ti 3 C 2 T X (a member of the MXene family) was used to immobilise enzyme sarcosine oxidase to fabricate a nanostructured biosensor. The device was applied for detection of sarcosine, a potential prostate cancer biomarker, in urine for the first time. The morphology and structures of MXene have been characterised by atomic force microscopy (AFM) and scanning electron microscopy (SEM). Electrochemical measurements, SEM and AFM analysis revealed that MXene interfaced with chitosan is an excellent support for enzyme immobilisation to fabricate a sensitive biosensor exhibiting a low detection limit of 18 nM and a linear range up to 7.8 µM. The proposed biosensing method also provides a short response time of 2 s and high recovery index of 102.6% for detection of sarcosine spiked into urine sample in a clinically relevant range., Competing Interests: Conflicts of Interest: The authors declare no conflict of interest.

Early diagnostics of diseases performed with minimal money and time consumption has become achievable due to recent advances in development of biosensors. These devices use biorecognition elements for selective interaction with an analyte and signal readout is obtained via different types of transducers. Operational characteristics of biosensors have been reported to improve substantially, when a diverse range of nanomaterials was employed. This review presents construction of electrochemical biosensors based on graphene, atomically thin 2D carbon crystals, which is currently intensively studied nanomaterial. The most attractive directions of graphene applications in biosensor preparation are discussed here including novel detection and amplification schemes exploiting graphene's unique electrochemical, physical and chemical properties. The future of graphene-based biosensors is most likely bright, but there is still a lot of work to do to fulfill high expectations.

Glycans are chains of carbohydrates attached to proteins (glycoproteins and proteoglycans) or lipids (glycolipids). Glycosylation is a posttranslational modification and glycans have a wide range of functions in a human body including involvement in oncological diseases. Change in a glycan structure cannot only indicate presence of a pathological process, but more importantly in some cases also its stage. Thus, a glycan analysis has a potential to be an effective and reliable tool in cancer diagnostics. Lectins are proteins responsible for natural biorecognition of glycans, even carbohydrate moieties still attached to proteins or whole cells can be recognized by lectins, what makes them an ideal candidate for designing label-free biosensors for glycan analysis. In this review we would like to summarize evidence that glycoprofiling of biomarkers by lectin-based biosensors can be really helpful in detecting prostate cancer.

Glycosylation of biomolecules is one of the most prevalent post- and co-translational modification in a human body, with more than half of all human proteins being glycosylated. Malignant transformation of cells influences glycosylation machinery resulting in subtle changes of the glycosylation pattern within the cell populations as a result of cancer. Thus, an altered terminal glycan motif on glycoproteins could provide a warning signal about disease development and progression and could be applied as a reliable biomarker in cancer diagnostics. Among all highly effective glycoprofiling tools, label-free electrochemical impedance spectroscopy (EIS)-based biosensors have emerged as especially suitable tool for point-of-care early-stage cancer detection. Herein, we highlight the current challenges in glycoprofiling of various cancer biomarkers by ultrasensitive impedimetric-based biosensors with low sample consumption, low cost fabrication and simple miniaturization. Additionally, this review provides a short introduction to the field of glycomics and lectinomics and gives a brief overview of glycan alterations in different types of cancer.

This is the first study showing pH dependence of three distinct redox sites within bilirubin oxidase (BOD) adsorbed on a nanocomposite modified electrode. The 1st redox centre with the highest redox potential Ec(1st)=404 mV vs. Ag/AgCl (614 mV vs. NHE at pH7.0) exhibited pH dependence with a slope -dEc(1st)/dpH=66(±3) mV under a non-turnover process. The 2nd redox centre with a potential Ec(2nd)=228 mV vs. Ag/AgCl (438 mV vs. NHE at pH7.0) was not dependent on pH in the absence and presence of O2. Finally, the 3rd redox site with a redox potential Ec(3rd)=92 mV vs. Ag/AgCl (302 mV vs. NHE at pH7.0) exhibited pH dependence for a cathodic process with -dEc(3rd)/dpH=70(±6) mV and for anodic process with -dEa(3rd)/dpH=73(±2) mV, respectively. Moreover, two break points for dependence of Ec(1st) or Ec(3rd) on pH were observed for the 1st (T1) site and the 3rd site assigned to involvement of two acidic amino acids (Asp105 and Glu463). A diagram of a potential difference between cathodic peaks of BOD as a dependence on pH is shown. The results obtained can be of interest for construction of biofuel cells based on BOD such as for generation of a low level of electricity from body fluids., (Copyright © 2013 Elsevier B.V. All rights reserved.)

We present here comparison of a build-up of two ultrasensitive lectin biosensors based on 2D or 3D architecture. A 2D lectin biosensor was prepared by a covalent immobilisation of lectin Sambucus nigra agglutinin (SNA) recognising sialic acid directly on a mixed self-assembled monolayer (SAM) on planar gold surfaces. A 3D biosensor was prepared by covalent immobilisation of SNA lectin on a mixed SAM layer formed on gold nanoparticles. Surface plasmon resonance technique allowed to follow kinetics of a mixed SAM (1:1 mixture of 11-mercaptoundecanoic acid and 6-mercaptohexanol) formation on a bare gold electrode and on an electrode modified by 5 nm and 20 nm gold nanoparticles (AuNPs). Results from the study revealed that a mixed SAM formation is slower on surfaces with increased curvature, the process of SAM formation on all surfaces is completed within 6 min, but a density of thiols on such surfaces differs significantly. Quartz crystal microbalance experiments showed that a surface density of immobilised lectin of (2.53 ± 0.01) pmol cm -2 was higher on planar gold surface compared to the surface modified by 20 nm AuNPs with a surface density of (0.94 ± 0.01) pmol cm -2 . Even though a larger amount of SNA lectin was immobilised on a surface of the 2D biosensor compared to the 3D biosensor, lectin molecules immobilised on AuNPs were more accessible for its analytes - glycoproteins fetuin and asialofetuin, containing different amount of sialic acid on the protein surface. Most likely a better accessibility of lectin for its analytes on a 3D surface and proper interfacial properties of a 3D surface are behind unprecedented detection limit down to aM level for the lectin biosensor based on such a nanoscale tuned interface.

This chapter would like to provide a short survey of the most promising concepts applied recently in analysis of glycoproteins based on lectins. The first part describes the most exciting analytical approaches used in the field of glycoprofiling based on integration of nanoparticles, nanowires, nanotubes, or nanochannels or using novel transducing platforms allowing to detect very low levels of glycoproteins in a label-free mode of operation. The second part describes application of recombinant lectins containing several tags applied for oriented and ordered immobilization of lectins. Besides already established concepts of glycoprofiling several novel aspects, which we think will be taken into account for future, more robust glycan analysis, are described including modified lectins, peptide lectin aptamers, and DNA aptamers with lectin-like specificity introduced by modified nucleotides. The last part of the chapter describes a novel concept of a glycocodon, which can lead to a better understanding of glycan-lectin interaction and for design of novel lectins with unknown specificities and/or better affinities toward glycan target or for rational design of peptide lectin aptamers or DNA aptamers.

Although membrane-bound dehydrogenases isolated from Gluconobacter sp. (mainly PQQ-dependent alcohol and fructose dehydrogenase) have been used for preparing diverse forms of bioelectronic interfaces for almost 2 decades, it is not an easy task to interpret an electrochemical behaviour correctly. Recent discoveries regarding redox properties of membrane-bound dehydrogenases along with extensive investigations of direct electron transfer (DET) or direct bioelectrocatalysis with these enzymes are summarized in this review. The main aim of this review is to draw general conclusions about possible electronic coupling paths of these enzymes on various interfaces via direct electron transfer or direct bioelectrocatalysis. A short overview of the metabolism and respiration chain in Gluconobacter relevant to interfacial electrochemistry is given. Biosensor devices based on DET or direct bioelectrocatalysis using membrane-bound dehydrogenases from Gluconobacter sp. are described briefly with the emphasis given on practical applications of preparing enzymatic biofuel cells. Moreover, interfacial electrochemistry of Gluconobacter oxydans related to the construction of microbial biofuel cells is also discussed.