Rapid nanocatalytic reaction using antibody-conjugated gold nanoparticles for simple and sensitive detection of parathyroid hormone

Abstract

Biomolecule-conjugated metal nanoparticles (NPs) have been primarily used as colorimetric labels in affinity-based bioassays for point-of-care testing. A facile electrochemical detection scheme using a rapid nanocatalytic reaction of a metal NP label is required to achieve more quantitative and sensitive point-of-care testing. Moreover, all the involved components should be stable in their dried form and solution. This study developed a stable component set that allows for rapid and simple nanocatalytic reactions combined with electrochemical detection and applied it for the sensitive detection of parathyroid hormone (PTH). The component set consists of an indium–tin oxide (ITO) electrode, ferrocenemethanol (FcMeOH), antibody-conjugated Au NPs, and ammonia borane (AB). Despite being a strong reducing agent, AB is selected because it is stable in its dried form and solution. The slow direct reaction between FcMeOH+ and AB provides a low electrochemical background, and the rapid nanocatalytic reaction allows for a high electrochemical signal. Under optimal conditions, PTH could be quantified in a wide range of concentrations in artificial serum, with a detection limit of ∼0.5 pg/mL. Clinical validation of the developed PTH immunosensor using real serum samples indicates that this novel electrochemical detection scheme is promising for quantitative and sensitive immunoassays for point-of-care testing.