用于比色和荧光检测中性粒细胞明胶酶相关脂褐素（NGAL）的银自动催化质子酶联免疫吸附测定法

IF 4.9 2区化学 Q1 CHEMISTRY, ANALYTICAL

Microchemical Journal Pub Date : 2024-09-01 DOI:10.1016/j.microc.2024.111551

Yu Liu, Yawen Liu, Fangfang Wang, Zhigang Zhang, Haiming Hu, Lei Xiong, Junping Zheng, Hongtao Liu

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引用次数: 0

摘要

传统的酶联免疫吸附试验（ELISA）在分析溃疡性结肠炎的潜在生物标志物--中性粒细胞明胶酶相关脂褐质（NGAL）时缺乏足够的灵敏度和准确性。在这里，我们开发了一种双读出质子酶联免疫吸附测定法，它对碱性磷酸酶水解和 O-苯二胺氧化进行级联放大。在这种新型的质子酶联免疫吸附试验中，碱性磷酸酶结合的抗体捕获样品中的 NGAL，碱性磷酸酶将 2-磷酸-L-抗坏血酸水解为抗坏血酸，然后抗坏血酸通过将银离子转化为银微丝阻断银催化的 O-苯二胺氧化。因此，NGAL 的浓度与发色反应和荧光反应的下降成正比。这种新型质子酶联免疫吸附分析仪的线性范围为 0.5-80 纳克/毫升，最低检测限低至 0.5 纳克/毫升。其灵敏度优于传统的酶联免疫吸附测定法和以往的研究。由于抗体与抗原的特异性相互作用和强大的级联放大作用，质子酶联免疫吸附试验具有良好的选择性。此外，质子酶联免疫吸附试验的双读取功能还具有方便性和兼容性等优点。总之，我们的研究为诊断溃疡性结肠炎提供了一种新型、灵敏的等离子体酶联免疫吸附测定。

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A silver auto-catalyzed plasmonic enzyme-linked immunosorbent assay for colorimetric and fluorescent detection of neutrophil gelatinase associated lipocalin (NGAL)

The conventional ELISA lacked sufficient sensitivity and accuracy for analyzing neutrophil gelatinase associated lipocalin (NGAL), a potential biomarker of ulcerative colitis. Here, we developed a dual-readout plasmonic ELISA with cascade amplification of alkaline phosphatase hydrolysis and O-phenylenediamine oxidation. In this novel plasmonic ELISA, alkaline phosphatase-conjugated antibodies capture NGAL from samples, alkaline phosphatase hydrolyzes 2-phospho-L-ascorbic acid into ascorbic acid, and then ascorbic acid blocks silver-catalyzed O-phenylenediamine oxidation by turning silver ions into silver microwires. Therefore, NGAL concentration is proportional to the decrease in chromogenic and fluorescent response. This novel plasmonic ELISA exhibited a linear range of 0.5–80 ng/mL, and the LOD was as low as 0.5 ng/mL. The sensitivity was superior to that of conventional ELISA and previous studies. The plasmonic ELISA exhibited favorable selectivity because of the specific antibody-antigen interaction and the robust cascade amplification. Besides, the dual-readout feature endowed the plasmonic ELISA with advantages in terms of convenience and compatibility. In summary, our study provided a novel and sensitive plasmonic ELISA for the diagnosis of ulcerative colitis.

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来源期刊

Microchemical Journal 化学-分析化学

CiteScore

8.70

自引率

8.30%

发文量

1131

审稿时长

1.9 months

期刊介绍： The Microchemical Journal is a peer reviewed journal devoted to all aspects and phases of analytical chemistry and chemical analysis. The Microchemical Journal publishes articles which are at the forefront of modern analytical chemistry and cover innovations in the techniques to the finest possible limits. This includes fundamental aspects, instrumentation, new developments, innovative and novel methods and applications including environmental and clinical field. Traditional classical analytical methods such as spectrophotometry and titrimetry as well as established instrumentation methods such as flame and graphite furnace atomic absorption spectrometry, gas chromatography, and modified glassy or carbon electrode electrochemical methods will be considered, provided they show significant improvements and novelty compared to the established methods.