Department of Chemistry and Chemistry Institute for Functional Materials, Pusan National University, Busan 46241, Republic of Korea.
EONE Laboratories, Incheon 22014, Republic of Korea.
Int J Biol Macromol. 2023 Jun 30;241:124574. doi: 10.1016/j.ijbiomac.2023.124574. Epub 2023 Apr 24.
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.
生物分子缀合的金属纳米粒子(NPs)主要用作基于亲和力的生物分析中的比色标记物,用于即时检测。为了实现更定量和更敏感的即时检测,需要使用金属 NP 标记物的快速纳米催化反应的简便电化学检测方案。此外,所有涉及的组件都应在干燥形式和溶液中稳定。本研究开发了一个稳定的组件集,允许快速简单的纳米催化反应与电化学检测相结合,并将其应用于甲状旁腺激素(PTH)的敏感检测。该组件集由氧化铟锡(ITO)电极、二茂铁甲醇(FcMeOH)、抗体偶联的 Au NPs 和氨硼烷(AB)组成。尽管 AB 是一种强还原剂,但由于其在干燥形式和溶液中稳定,因此被选择使用。FcMeOH 和 AB 之间的缓慢直接反应提供了低的电化学背景,而快速的纳米催化反应则产生了高的电化学信号。在最佳条件下,PTH 可以在人工血清的宽浓度范围内进行定量,检测限约为 0.5 pg/mL。使用实际血清样本对开发的 PTH 免疫传感器进行临床验证表明,这种新型电化学检测方案有望用于即时检测的定量和敏感免疫分析。