The objective of this paper is development of an inexpensive point-of-care sensor for detecting the primary heart failure marker peptide, NT-proBNP. The device technology is based on an antibody sandwich assay, but with three innovative aspects. First, chemical amplification is carried out via oxidation of silver nanoparticles (NPs) attached to signaling antibodies rather than by enzymatic amplification. The electrochemical method is faster and eliminates the need for long-term storage of enzymes. Second, the antibody sandwich is formed on mobile magnetic beads. This enhances the rate of mass transfer of the biomarker and the signaling antibody to the primary detection antibody, which is immobilized on the magnetic beads. Third, the sensor itself is fabricated on a paper platform with screen-printed electrodes. This coupled with assembly by simple paper folding, keeps the cost of the sensor low. Here, we report on two separate assays. The first is based on a simple biotin-streptavidin conjugate, which is a preliminary model for the antibody assay. The results indicate a detection limit of 2.1 pM of silver NPs and an assay time of 7 min. The actual NT-proBNP antibody assay takes somewhat longer, and the dynamic detection range is higher: 2.9–582 nM. On the basis of the results presented in this paper, we conclude that this inexpensive paper-based sensor represents a viable technology for point-of-care testing of NT-proBNP, but nevertheless several challenges remain prior to clinical implementation. These include attaining a lower detection limit and better reproducibility, and optimizing the device for human blood.
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