In this study, a signal-on nanobiosensor based on bivalent aptamer-Cu nanocluster was designed and optimized for specific and sensitive detection of VEGF165. The VEGF165 is known as a promising biomarker in different diseases such as cancer in the angiogenic stage. Detection and quantification of VEGF165 is a crucial step in diagnosis and monitoring the treatment plan. The represented nanostructure consists of multimerized VEGF165 aptamer joint with ssDNA based linker in the middle and poly thymine sequences on both 3' and 5' ends as a template for Cu-nanocluster supraparticle formation. This self-assembled structure leads to accurate controlling of aggregation in the presence of VEGF165. This study is the first report for Cu nanocluster nucleation on ploy thymine tails of ssDNA which performed in two reduction steps to form stable CuNC supraparticle. The sensing strategy was designed based on the target-induced structure switching mode of the aptamer. In the presence of VEGF165, due to self-assembly induced emission and aggregation-induced emission phenomena this nanostructure depicted the visible wavelength shift and enhancement in the fluorescence emission intensity. Also, the results of the analytical performance of this nanobiosensor indicated the LOD of 12 pM which revealed high rate sensitivity. This aptasensor exhibited stability and decent response linearity range (10-800 pM, R2 = 0.9943). The selectivity and specificity assessment showed high discriminant capability in the real serum sample. In conclusion, this signal-on nanobiosensor provides a facile, sensitive and reliable assay for clinical monitoring of the VEGF165 concentration in serum without further sample preparation.
Keywords: Aggregation-induced emission; Bivalent aptamer; Cu nanocluster; Nanobiosensor; Target-induced structure switching mode; VEGF(165).
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