Highly sensitive detection of human cardiac myoglobin using a reverse sandwich immunoassay with a gold nanoparticle-enhanced surface plasmon resonance biosensor

被引：91

作者：

Gnedenko, Oksana V. ^{[1
]}

Mezentsev, Yury V. ^{[1
]}

Molnar, Andrey A. ^{[1
]}

Lisitsa, Andrey V. ^{[1
]}

Ivanov, Alexis S. ^{[1
]}

Archakov, Alexander I. ^{[1
]}

机构：

[1] IBMC RAMS, Moscow 119121, Russia

来源：

ANALYTICA CHIMICA ACTA | 2013年 / 759卷

关键词：

Surface plasmon resonance; Reverse sandwich immunoassay; Cardiac myoglobin; Gold nanoparticles; Signal amplification; ACUTE MYOCARDIAL-INFARCTION; PROSTATE-SPECIFIC ANTIGEN; BIOCHEMICAL MARKERS; SPR; ANTIBODIES; DIAGNOSIS; ASSAYS; ELISA; AMPLIFICATION; TECHNOLOGY;

D O I：

10.1016/j.aca.2012.10.053

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A highly sensitive reverse sandwich immunoassay for the detection of human cardiac myoglobin (cMb) in serum was designed utilizing a gold nanoparticle (AuNP)-enhanced surface plasmon resonance (SPR) biosensor. First, a monoclonal anti-cMb antibody (Mab1) was covalently immobilized on the sensor surface. AuNPs were covalently conjugated to the second monoclonal anti-cMb antibody (Mab2) to form an immuno-gold reagent (Mab2-AuNP). The reverse sandwich immunoassay consists of two steps: (I) mixing the serum sample with Mab2-AuNP and incubation for the formation of cMb/Mab2-AuNP complexes and (2) sample injection over the sensor surface and evaluation of the Mab1/cMb/Mab2-AuNP complex formation, with the subsequent calculation of the cMb concentration in the serum. The biosensor signal was amplified approximately 30-fold compared with the direct reaction of cMb with Mab1 on the sensor surface. The limit of detection of cMb in a human blood serum sample was found to be as low as 10 pM (approx. 0.18 ngmL(-1)), and the inter-assay coefficient of variation was less than 3%. Thus, the developed SPR-based reverse sandwich immunoassay has a sensitivity that is sufficient to measure cMb across a wide range of normal and pathological concentrations, allowing an adequate estimation of the disease severity and the monitoring of treatment. (C) 2012 Elsevier B.V. All rights reserved.

引用

页码：105 / 109

页数：5

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