Reports have indicated that warfarin is the most widely prescribed anticoagulant. However, traditionally prescribed doses for each patient may be too low or too high. The therapeutic effect is often hindered by a lack of evidence-based medical information. Herein, our aim is to provide this information. To accomplish this challenge, we report the development of a novel assay based on biotinylated tetrahedral DNA as a capture probe and fullerene (C60)-based nanomaterial as a redox probe using an ultrasensitivity assay with the Vitamin K epoxide reductase complex, subunit 1 (VKORC1). Platinum porous nanoparticles (PtPNPs) were modified on amino-terminated polyamidoamine (PAMAM)-functionalized C60 nanoparticles (C60NPs). The resultant C60NPs-PAMAM-PtPNPs were used as a redox probe. In this design, C60 exhibited excellent redox activity that was triggered by tetraoctylammonium bromide (TOAB). To improve the immobilization of the tetrahedral DNA capture probe, avidin was introduced during the fabrication of the biosensor because it can provide more active sites for the immobilization capture probe. The free-standing probe on top of the tetrahedral DNA served as a receptor to hybridize with target DNA directly. Different pulse voltammetry (DPV) was applied to record the electrochemical signals, which increased linearly with the target DNA. Under optimal conditions, the prepared biosensor showed a wide linear relationship, from 1 pM to 10 nM, with detection limits of 0.33 pM. This strategy demonstrates a new avenue for the determination of tumour-related mutated nucleotides in biosamples.
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