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首页> 外文期刊>RSC Advances >Ternary covalent conjugate (antibody-gold nanoparticle-peroxidase) for signal enhancement in enzyme immunoassay
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Ternary covalent conjugate (antibody-gold nanoparticle-peroxidase) for signal enhancement in enzyme immunoassay

机译：三元共价结合物（抗体-金纳米粒子-过氧化物酶），用于酶免疫测定中的信号增强

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摘要

A synthesis of a new horseradish peroxidase (HRP) conjugate and antibody covalently bound with gold nanoparticles (GNPs) with a high enzyme content was first developed. The synthesis conditions were optimized by varying the concentrations of the reacting compounds. In the conjugate, which was synthesized under the optimized conditions, one gold nanoparticle carrier ((sic) 30 nm) contained 13 active HRP and 1 anti-troponin I antibody molecule at its surface. The applicability of the proposed ternary conjugate was successfully proved in chemiluminescent ELISA by determining human troponin I. The sensitivity of the assay using the ternary conjugate was higher than that of the assay with the streptavidin-biotin system (biotinylated antibody and streptavidin-HRP conjugate). High sensitivity was achieved due to the dual signal amplification from use of a potent enhancer of HRP-catalyzed chemiluminescence (3-(10'-phenothiazinyl) propane-1-sulfonate) and the high HRP content in the conjugate. A replacement of two-step streptavidin-biotin amplification system with the ternary conjugate decreased the assay time. The use of the proposed conjugate significantly reduced antibody and HRP consumption per assay.
机译：首先开发了一种新的辣根过氧化物酶（HRP）共轭物和与具有高酶含量的金纳米颗粒（GNP）共价结合的抗体的合成方法。通过改变反应化合物的浓度来优化合成条件。在优化条件下合成的缀合物中，一种金纳米颗粒载体（约30 nm）在其表面含有13个活性HRP和1个抗肌钙蛋白I抗体分子。通过测定人肌钙蛋白I在化学发光ELISA中成功证明了所提出的三价结合物的适用性。使用三价结合物的测定灵敏度高于抗生蛋白链菌素-生物素系统（生物素化抗体和抗生蛋白链菌素-HRP结合物）的灵敏度。 。由于使用了有效的HRP催化化学发光增强剂（3-（10'-吩噻嗪基）丙烷-1-磺酸盐）增强了双重信号，并且结合物中的HRP含量较高，因此实现了高灵敏度。用三元偶联物替代两步链霉亲和素-生物素扩增系统可减少测定时间。所提出的缀合物的使用显着降低了每次测定的抗体和HRP消耗。

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