4.7 Article

Disposable immunoplatforms for the simultaneous determination of biomarkers for neurodegenerative disorders using poly(amidoamine) dendrimer/gold nanoparticle nanocomposite

Journal

ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 413, Issue 3, Pages 799-811

Publisher

SPRINGER HEIDELBERG
DOI: 10.1007/s00216-020-02724-3

Keywords

Dual immunosensing platform; Tau; TDP-43; Amperometric detection; Plasma; Brain tissue extracts

Funding

  1. Spanish Ministerio de Economia y Competitividad [CTQ2015-64402-C21-R, BES-2016076606]
  2. Ministerio de Ciencia, Innovacion y Universidades [RTI2018096135-B-I00]
  3. TRANSNANOAVANSENS-CM Program from the Comunidad deMadrid [S2018/NMT-4349]
  4. FAPESP [2018/14130-7]
  5. AES-ISCIII program [PI17CIII/00045]
  6. Fundacion Tatiana Perez de Guzman el Bueno
  7. FPU - Spanish Ministerio de Educacion, Cultura y Deporte

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This study developed a disposable immunoplatform for the simultaneous determination of two protein biomarkers involved in neurodegenerative disorders. The method exhibited high sensitivity and selectivity for detecting the target proteins, allowing for quick analysis in raw samples.
Early diagnosis in primary care settings can increase access to therapies and their efficiency as well as reduce health care costs. In this context, we report in this paper the development of a disposable immunoplatform for the rapid and simultaneous determination of two protein biomarkers recently reported to be involved in the pathological process of neurodegenerative disorders (NDD), tau protein (tau), and TAR DNA-binding protein 43 (TDP-43). The methodology involves implementation of a sandwich-type immunoassay on the surface of dual screen-printed carbon electrodes (dSPCEs) electrochemically grafted with p-aminobenzoic acid (p-ABA), which allows the covalent immobilization of a gold nanoparticle-poly(amidoamine) (PAMAM) dendrimer nanocomposite (3D-Au-PAMAM). This scaffold was employed for the immobilization of the capture antibodies (CAbs). Detector antibodies labeled with horseradish peroxidase (HRP) and amperometric detection at - 0.20 V (vs. Ag pseudo-reference electrode) using the H2O2/hydroquinone (HQ) system were used. The developed methodology exhibits high sensitivity and selectivity for determining the target proteins, with detection limits of 2.3 and 12.8 pg mL(-1) for tau and TDP-43, respectively. The simultaneous determination of tau and TDP-43 was accomplished in raw plasma samples and brain tissue extracts from healthy individuals and NDD-diagnosed patients. The analysis can be performed in just 1 h using a simple one-step assay protocol and small sample amounts (5 mu L plasma and 2.5 mu g brain tissue extracts).

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