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.
BACKGROUND: A sensitive nanostructured immunoelectrode based on poly(allylamine) (PAH) sandwich is developed for nonstructural 1 (NS1) of dengue virus. NS1 is a secretory protein abundant in the acute phase of disease associated to hemorrhagic fever. Anti-NS1 antibodies are immobilized on the electrode surface by a thin layer of PAH assembled on carboxylated carbon nanotubes (CNTs). PAH is cationic polymer acting as bi-functional agent to tightly attach CNTs to the electrode surface and anti-NS1 antibodies through their Fc terminal, avoiding random immobilization. Electrochemical responses of immunoassay are generated at a controlled potential by a reaction between H 2 O 2 and peroxidase enzyme conjugated to anti-NS1 antibodies.
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