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1.
J Mater Chem B ; 12(26): 6342-6350, 2024 Jul 03.
Article in English | MEDLINE | ID: mdl-38856318

ABSTRACT

The enzyme-linked immunosorbent assay (ELISA) remains the prevailing method for quantifying protein biomarkers. Enzymatic signal generation and amplification are key mechanisms that govern its analytical performance. This study reports the synthesis and application of microscale metal-organic framework (MOF)/enzyme composite particles as a novel detection probe to substantially enhance the sensitivity of ELISA. An optimal one-pot approach was established to incorporate a substantial amount of streptavidin-horseradish peroxidase (SA-HRP) either within or on the surface of the metal-azolate framework (MAF-7) microparticles. This approach enables the labeling of a single sandwich antibody-antigen complex with numerous enzymes, which markedly amplifies the enzymatic colorimetric signal generation. Moreover, MAF-7 caging was found to enhance the reactivity of the caged HRP enzyme, further promoting the overall detection sensitivity of ELISA. Compared to other developments that are often associated with more complicated detection modalities, our method is compatible with standard immunoassays and commonly used photometrical signal detection. The implementation of this strategy in the detection of CD147 results in a remarkably low limit of detection of 2.8 fg mL-1, representing a 105-fold improvement compared to that obtained with the standard ELISA. Moreover, the heightened sensitivity of this technique renders it particularly suitable for diagnosing breast cancer, thus presenting a promising tool for the early detection of the disease in clinical settings.


Subject(s)
Enzyme-Linked Immunosorbent Assay , Extracellular Vesicles , Metal-Organic Frameworks , Metal-Organic Frameworks/chemistry , Humans , Extracellular Vesicles/chemistry , Horseradish Peroxidase/chemistry , Horseradish Peroxidase/metabolism , Particle Size , Surface Properties , Biomarkers, Tumor/analysis , Biomarkers/analysis , Limit of Detection
2.
Small ; : e2002517, 2020 Oct 08.
Article in English | MEDLINE | ID: mdl-33090659

ABSTRACT

Controlled deposition of 2D multilayered nanomaterials onto different electrodes to design a highly sensitive biosensing platform utilizing their active inherent electrochemistry is extremely challenging. Herein, a green, facile, and cost-effective one-pot deposition mechanism of 2D MXene-Ti3C2Tx nanosheets (MXNSs) onto conductive electrodes within few minutes via electroplating (termed electroMXenition) is reported for the first time. The redox reaction in the colloidal MXNS solution under the effect of a constant applied potential generates an electric field, which drives the nanoparticles toward a specific electrode interface such that they are cathodically electroplated. A task-specific ionic liquid, that is, 4-amino-1-(4-formyl-benzyl) pyridinium bromide (AFBPB), is exploited as a multiplex host arena for the substantial immobilization of MXNSs and covalent binding of antibodies. A miniaturized, single-masked gold dual interdigitated microelectrode (DIDµE) is microfabricated and presented by investigating the benefit of AFBPB coated on MXNSs. The resulting MXNSs-AFBPB-film-modified DIDµE biosensor exhibited a 7× higher redox current than bare electrodes owing to the uniform deposition. Using Apo-A1 and NMP 22 as model bladder cancer analytes, this newly developed dual immunosensor demonstrated precise and large linear ranges over five orders of significance with limit of detection values as low as 0.3 and 0.7 pg mL-1, respectively.

3.
Biosens Bioelectron ; 160: 112220, 2020 Jul 15.
Article in English | MEDLINE | ID: mdl-32339151

ABSTRACT

The patterned LIG flakes are generally not interconnected due to the line gap of the laser ray, leading to lower uniform conductivity and fragile graphene. Thus, the fabrication of a highly conductive and mechanically robust LIG-based biosensing platform remains challenging. In this study, the fabrication of a flexible electrochemical biosensor is reported based on poly (3, 4-ethylene dioxythiophene)-poly (styrene sulfonate) (PEDOT:PSS) modified 3-dimensional (3D) stable porous laser-induced graphene (LIG) for the detection of glucose and pH. PEDOT:PSS was spray-coated on the LIG to improve electrode robustness and deliver uniform electrical conductivity. The as-prepared PEDOT:PSS modified LIG (PP/LIG) was characterized using field-emission scanning electron microscopy (FESEM), x-ray photoelectron spectroscopy (XPS), Raman spectroscopy, and Fourier-transform infrared spectroscopy (FTIR). Platinum and palladium nanoparticles (Pt@Pd) were successfully electrodeposited on PP/LIG, markedly enhancing the electrocatalytic activity for glucose detection. The fabricated biosensor exhibited an excellent amperometric response to glucose with a wide linear range of 10 µM - 9.2 mM, a high sensitivity of 247.3 µAmM-1cm-2, and a low detection limit (LOD) of 3 µM, with high selectivity. In addition, the pH sensor was functionalized by the polyaniline (PANI) on PP/LIG, and it also exhibited excellent potentiometric response with a high sensitivity of 75.06 mV/pH in the linear range of pH 4 - 7. Ultimately, the feasibility of the biosensor was confirmed by the analysis of human perspiration collected during physical exercise. This approach validates the utility of the novel fabrication procedure, and the potential of the LIG-conductive polymer composite for biosensing applications.


Subject(s)
Glucose/analysis , Graphite/chemistry , Polystyrenes/chemistry , Sweat/chemistry , Thiophenes/chemistry , Biosensing Techniques/methods , Electric Conductivity , Electrochemical Techniques/methods , Humans , Hydrogen-Ion Concentration , Limit of Detection , Porosity
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