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1.
Mikrochim Acta ; 188(10): 334, 2021 Sep 08.
Article in English | MEDLINE | ID: mdl-34498145

ABSTRACT

An innovative sensing assay is described for point-of-care (PoC) quantification of a biomarker of Alzheimer's disease, amyloid ß-42 (Aß-42). This device is based on a cellulose paper-dye test strip platform in which the corresponding detection layer is integrated by applying a molecularly imprinted polymer (MIP) to the cellulose paper surface. Briefly, the cellulose paper is chemically modified with a silane to subsequently apply the MIP detection layer. The imprinting process is confirmed by the parallel preparation of a control material, namely a non-imprinted polymer (NIP). The chemical changes of the surface were evaluated by Fourier transform infrared spectroscopy (FTIR), contact angle, and thermogravimetric analysis (TG). Proteins and peptides can be quantified by conventional staining methods. For this purpose, Coomassie blue (CB) was used as a staining dye for the detection and quantification of Aß-42. Quantitative determination is made possible by taking a photograph and applying an appropriate mathematical treatment to the color coordinates provided by the ImageJ program. The MIP shows a linear range between 1.0 ng/mL and 10 µg/mL and a detection limit of 0.71 ng/mL. Overall, this cellulose-based assay is suitable for the detection of peptides or proteins in a sample by visual comparison of color change. The test strip provides a simple, instrument-free, and cost-effective method with high chemical stability, capable of detecting very small amounts of peptides or proteins in a sample, and can be used for the detection of any (bio)molecule of interest.


Subject(s)
Amyloid beta-Peptides/blood , Cellulose/chemistry , Colorimetry/methods , Immunoassay/methods , Peptide Fragments/blood , Amyloid beta-Peptides/chemistry , Amyloid beta-Peptides/immunology , Animals , Antibodies, Immobilized/immunology , Biomarkers/blood , Cattle , Colorimetry/instrumentation , Coloring Agents/chemistry , Immunoassay/instrumentation , Limit of Detection , Molecularly Imprinted Polymers/chemistry , Peptide Fragments/chemistry , Peptide Fragments/immunology , Point-of-Care Testing , Rosaniline Dyes/chemistry
2.
Biosens Bioelectron ; 77: 978-85, 2016 Mar 15.
Article in English | MEDLINE | ID: mdl-26544873

ABSTRACT

A nano-molecularly imprinted polymer (N-MIP) assembled on a screen-printed electrode for the cardiac troponin T (cTnT) was developed. The biomimetic surface was obtained by a co-polymer matrix assembled on the reduced graphene oxide (RGO) electrode surface. The cTnT active sites were engineered using pyrrole and carboxylated pyrrole that was one-step electropolymerized jointly with cTnT by cyclic voltammetry. The stepwise preparation of the biomimetic surface was characterized by cyclic and differential pulse voltammetries using the ferrocyanide/ferricyanide as redox probe. Structural and morphological characterization was also performed. The optimal relation of pyrrole and pyrrole-3-acid carboxylic to perform the cTnT biomimetic nanosurface was obtained at 1:5 ratio. The analytical performance of cTnT N-MIP performed by differential pulse voltammetry showed a linear range from 0.01 to 0.1 ngmL(-1) (r=0.995, p«0.01), with a very low limit of detection (0.006 ngmL(-1)). The synergic effect of conductive polymer and graphene forming 3D structures of reactive sites resulted in a N-MIP with excellent affinity to cTnT binding (KD=7.3 10(-13) molL(-1)). The N-MIP proposed is based on a simple method of antibody obtaining with a large potential for point-of-care testing applications.


Subject(s)
Conductometry/instrumentation , Electrodes , Graphite/chemistry , Immunoassay/instrumentation , Molecular Imprinting/methods , Troponin T/blood , Electric Conductivity , Electroplating/methods , Equipment Design , Equipment Failure Analysis , Humans , Photography/methods , Reproducibility of Results , Sensitivity and Specificity
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