Exploring various carbon nanomaterials-based electrodes modified with polymelamine for the reagentless electrochemical immunosensing of Claudin18.2.

Claudin18.2 (CLDN18.2) is a tight junction protein often overexpressed in various solid tumors, including gastrointestinal and esophageal cancers, serving as a promising target and potential biomarker for tumor diagnosis, treatment assessment, and prognosis. Despite its significance, no biosensor has been reported to date for the detection of CLDN18.2. Here, we present the inaugural immunosensor for CLDN18.2. In this study, an amine-rich conducting polymer of polymelamine (PM) was electrografted onto different carbon nanomaterial-based screen-printed electrodes (SPEs), including carbon (C), graphene (Gr), graphene oxide (GO), carbon nanotube (CNT), and carbon nanofiber (CNF) via cyclic voltammetry. A comparative study was performed to explore the best material for the preparation of the PM-modified electrodes to be used as in-situ redox substrate for the immunosensor fabrication. The surface chemistry and structural features of pristine and PM-deposited electrodes were analyzed using Raman and scanning electron microscopy (SEM) techniques. Our results showed that the PM deposited on Gr and CNT/SPEs exhibited the most significant and stable redox behavior in PBS buffer. The terminal amine moieties on the PM-modified electrode surfaces were utilized for immobilizing anti-CLDN18.2 monoclonal antibodies via N-ethyl-N'-(3-(dimethylamino)propyl)carbodiimide/N-hydroxysuccinimide chemistry to construct the electrochemical immunosensor platform. Differential pulse voltammetry-based immunosensing of CLDN18.2 protein on BSA/anti-CLDN18.2/PM-Gr/SPE and BSA/anti-CLDN18.2/PM-CNT/SPE exhibited excellent selectivity against other proteins such as CD1, PDCD1, and ErBb2. The limits of detection of these two immunosensor platforms were calculated to be 7.9 pg/mL and 0.104 ng/mL for the CNT and Gr immunosensors, respectively. This study demonstrated that the PM-modified Gr and CNT electrodes offer promising platforms not only for the reagentless signaling but also for covalent immobilization of biomolecules. Moreover, these platforms offer excellent sensitivity and selectivity for the detection of CLDN18.2 due to its enhanced stable redox activity. The immunosensor demonstrated promising results for the sensitive detection of CLDN18.2 in biological samples, addressing the critical need for early gastric cancer diagnosis.

Redox probe-free electrochemical immunosensor utilizing electropolymerized melamine on reduced graphene oxide for the point-of-care diagnosis of gastric cancer.

Pepsinogen I (PG I) is a biomarker that plays a crucial role in the diagnosis of gastric cancer. The development of biosensor to monitor PG I overexpression in serum is crucial for early gastric cancer diagnosis, offering a less invasive alternative to the costly and uncomfortable gastroscopy procedure. This study presents a cost-efficient, scalable and disposable label-free biosensing strategy for detecting PG I, utilizing a redox-active polymelamine electrodeposited on a reduced graphene oxide screen-printed electrode surface (PM-rGO/SPE). Under optimized conditions, the conducting polymer PM was deposited on the rGO/SPE via a potentiodynamic method. The structural and morphological features of PM-rGO/SPE were analyzed with the assistance of Raman and Scanning Electron Microscopy analysis. Specific monoclonal anti-PG I antibodies were immobilized on the in situ prepared redox-active layer via EDC/NHS chemistry to develop a novel electrochemical immunosensor. Unlike the traditional immunosensing strategies which utilizes external redox probe solution for measuring the signal, the developed configuration allowed for redox-probe free monitoring of current changes of the redox active PM resulting from the formation of the immunocomplex on the electrode surface. Utilizing this method, PG I detection spanned a clinically relevant concentration range of 0.01-200 ng/mL, with a low limit of detection at 9.1 pg/mL. The electrochemical immunosensor demonstrated specificity against other biomarkers such as PDCD1, ErBb2, and CD28 with negligible interference. The immunosensor exhibited excellent recovery capabilities for PG I detection in serum samples. These findings underscore the potential of the PM-rGO/SPE immunosensor as a point-of-care diagnostic tool for gastric cancer.

Label-free genosensing of dengue serotypes with an electrodeposited reduced graphene oxide-tris(bipyridine)ruthenium(II).

Constructing a label-free electrochemical transducer platform without compromising inherent biocompatibility against specific bioreceptor remains challenging, particularly probing nucleic acid hybridization at electrode interface without external redox-mediator. Here, we show that electrochemically reduced graphene oxide-tris(bipyridine)ruthenium(II) (ErGO-TBR) nanosheets electrodeposited on carbon screen printed electrode can quantify hybridization of clinically important target sequences specific to serotypes of dengue virus (DENV) non-structural 1 (NS1) protein. Different variables including deposition potential, time, and electrolytic composition were optimized for fabrication of label-free transducer platform. Structural and electrochemical properties of ErGO-TBR/SPE were comprehensively elucidated using microscopic and spectroscopic techniques. Electrochemical quartz crystal microbalance (EQCM) analysis reveals the growth of electrodeposited redox-active species on the electrode interface. Surface functional group investigations suggested that TBR deposited on the basal and edges of ErGO substrate via electrostatic and π-π interactions. Functionalization of bio-affinity layer (B) on ErGO-TBR/SPE enables better loading of probe DNA (PDNA) toward specific detection of DENV target DNA (TDNA) with an ultralow detection limit promising for clinical diagnosis. Scalable chronoamperometry-based redox-active surface growth, customizable bioactivation strategy and external mediator-less probing of nucleic acid hybridization make the present system suitable for other translational application in healthcare diagnosis.

Electropolymerized Melamine on Electrochemically Reduced Graphene Oxide: Growth Mechanistics, Electrode Processing, and Amperometric Sensing of Acyclovir.

Metal-free, cost-efficient, redox-active electrode materials, combining graphene derivatives with nitrogen-rich polymelamine (PM), are widely explored as an interface layer for electrocatalysis and an electrochemical sensor platform. However, conventional chemical routes often yield derivatives of PM suffering from impaired redox behavior, restricting their electron-transfer kinetics. Herein, an optimal potentiodynamic method has been established to electrodeposit PM on electrochemically reduced graphene oxide (ErGO). A supporting electrolyte, containing Cl-, enhances the formation of intermediates NH3+ and ═NH2+ at the monomeric melamine, eventually interacting with the residual oxygenated functional groups of ErGO to form PM. In situ Raman spectrum analysis revealed the influence of the defective area and the graphitization ratio on the ErGO surface during the course of electropolymerization of melamine. Under optimal electrodeposition conditions (E = 0-1.6 V; ν = 0.1 V/s), the amount of electrodeposited PM on the ErGO surface was determined to be 16.5 µg/(cycle·cm2), using electrochemical quartz crystal microbalance analysis. An ErGO-PM-modified glassy carbon electrode (GCE) and a screen-printed electrode exhibit the direct electrooxidation of acyclovir (ACV). Amperometric analyses of ErGO-PM-modified electrodes exhibited the lowest detection limit of 137.4 pM with analytical robustness, rapid steady state, and reproducibility promising for ACV detection in complex biological matrices.

Electrochemical and DFT studies of andrographolide on electrochemically reduced graphene oxide for anti-viral herbaceutical sensor.

Herbal extracts are re-emerging as potential remedies for various vector-borne diseases. Amongst several phytochemicals, active ingredients of Andrographis paniculata extract is regarded as promising for dengue fever, caused by Aedes species. However, fingerprinting the active phytochemicals from herbal extracts are often relies on sophisticated analytical techniques which are not universally accessible. Herein, an electrochemically reduced graphene oxide on glassy carbon electrode (ErGO/GCE) has been devised as user-friendly and cost-effective sensor platform for fingerprinting of andrographolide (AG) in anti-dengue polyherbal formulation, i.e., Nilavembu kudineer powder. Confocal laser Raman and X-ray photoelectron spectral analyses revealed that the ErGO surfaces exert structural defects augmenting the conductivity at the electrode interface. DFT investigations enabled that C-3 and C-18 OH groups in AG is involved in the electrooxidation and adsorption-diffusion at the ErGO interface, respectively. Complementary electrochemical studies revealed that the diffusion-controlled process follows 1e-/1H+ transfer. Under optimal experimental conditions, ErGO sensor platform exhibit an amplified current sensitivity of 13.3 µA µM-1. cm-2 in the studied analyte concentration range of 10-400 µM. From the polyherbal extract and clinical sample analysis, the proposed sensor system offers selective, and sensitive detection of target AG regardless of common interferents.

Opto-electrochemical functionality of Ru(II)-reinforced graphene oxide nanosheets for immunosensing of dengue virus non-structural 1 protein.

Transforming the structural and functional properties of carbon nanostructures are highly beneficial for healthcare diagnostics. This research demonstrates the functionalization of opto-electrochemically active ruthenium bipyridine complex (Ru(II)) on the surface of graphene oxide (GO), enabling a dual-functional immunoprobe for the detection of non-structural 1 (NS1) protein, a dengue biomarker. Structural investigations reveals that Ru(II) has intermolecular bonding with functional groups of GO. Ultraviolet photoelectron spectral readouts display the changes in the work function and ionization energy of GO, supporting the functionalization of Ru(II). Bio-affinity layers of protein-G (Pro-G) at GO-Ru(II) electrode interface promotes the localization of monoclonal antibodies (mAb) selective for binding the epitopes of NS1 antigen. The chronoamperometric and fluorescence quenching-based immunoassays showed a linear response with a lowest detection limit of 0.38 and 0.48 ng/mL, respectively. Under optimal condition, the developed immunosensor studied to have retained stability/sensitivity toward NS1 without impact from interferents. The dual functional immuno-bioprobe translated from GO-Ru(II) conjugated nanostructures offers new insights for further studies in on-site diagnosis.