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首页> 外文期刊>Science of advanced materials >Development of Colorimetric Enzyme-Ball for Signal Amplification of Enzyme-Linked Immunosorbent Assay
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Development of Colorimetric Enzyme-Ball for Signal Amplification of Enzyme-Linked Immunosorbent Assay

机译：比色酶球用于酶联免疫吸附测定信号放大的开发

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摘要

Enzyme-linked immunosorbent assay (ELISA) is one of the most conventional protein assay methods and a well-established technique that offers multiple advantages such as selectivity, rapidity, and simplicity. However, colorimetric assay methods normally have lower sensitivity (i.e., picomolar concentrations) than other assay techniques. Bovine serum albumin (BSA) nanoparticles are able to encapsulate various biomaterials such as proteins and genes, as well as chemotherapeutic agents. In this study, we developed a novel colorimetric nanoprobe, enzyme-balls consisting of BSA and horseradish peroxidase (HRP), for signal amplification in ELISA. The colorimetric HRP enzyme-ball was synthesized by a desolvation technique using a mixed solution of BSA and HRP. It was then modified with streptavidin and a targeting antibody for the detection of the targeted protein. It has replaced the HRP-labeled antibody as a signaling probe for use in conventional ELISA. The localized, encapsulated HRP enzymes in the BSA nanoball were used to amplify the signal of the target probe. Prostate specific antigen (PSA), used as a model protein, was captured on the plate by the immobilized capturing antibody through the immune reaction that resulted from the immunoreaction, and the enzyme-ball was attached to the PSA-positive region. For the detection of PSA, we used the reaction of HRP-encapsulated enzyme-balls with tetramethylbenzidine (TMB), a substrate of HRP. We then stopped the reaction with 2 M H_2SO_4. The resulting products were analyzed by ultraviolet-visible (UV-vis) spectroscopy at 450 nm. This novel nanoprobe enabled us to detect the targeted protein at femtomolar concentrations in a short period. This method, based on the use of HRP-conjugated antibodies, is very simple and highly sensitive for use in established ELISA techniques and can be used for detecting specific protein markers associated with tumors, bacterial infections, or other diseases.
机译：酶联免疫吸附测定（ELISA）是最常规的蛋白质测定方法之一，是一种成熟的技术，具有多种优势，例如选择性，快速和简便。然而，比色测定法通常比其他测定技术具有较低的灵敏度（即皮摩尔浓度）。牛血清白蛋白（BSA）纳米颗粒能够封装各种生物材料，例如蛋白质和基因，以及化学治疗剂。在这项研究中，我们开发了一种新型比色纳米探针，由BSA和辣根过氧化物酶（HRP）组成的酶球，用于ELISA中的信号放大。使用BSA和HRP的混合溶液通过去溶剂化技术合成比色HRP酶球。然后用抗生蛋白链菌素和靶向抗体对其进行修饰，以检测目标蛋白。它已取代了HRP标记的抗体，作为常规ELISA中使用的信号探针。 BSA纳米球中的定位，封装HRP酶用于扩增目标探针的信号。用作模型蛋白的前列腺特异性抗原（PSA）被固定的捕获抗体通过免疫反应引起的免疫反应捕获在板上，并且酶球附着在PSA阳性区域。为了检测PSA，我们使用了HRP封装的酶球与HRP的底物四甲基联苯胺（TMB）的反应。然后，我们用2 M H_2SO_4终止反应。通过在450nm下的紫外-可见（UV-vis）光谱分析所得产物。这种新颖的纳米探针使我们能够在短时间内检测到飞摩尔浓度的目标蛋白质。该方法基于使用HRP偶联的抗体，非常简单并且对建立的ELISA技术非常敏感，可用于检测与肿瘤，细菌感染或其他疾病相关的特定蛋白质标记。

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  • 来源
    《Science of advanced materials》 |2014年第11期|2572-2576|共5页
  • 作者

    Jin-Ha Choi; Yong Taik Lim; Byung-Keun Oh;

  • 作者单位

    Department of Chemical and Biomolecular Engineering, Sogang University, 35 Baekbeom-Ro, Mapo-Gu, Seoul, 121-742, Republic of Korea;

    SKKU Advanced Institute of Nanotechnology (SAINT), School of Chemical Engineering, Sungkyunkwan University, Suwon, 440-746, Republic of Korea;

    Department of Chemical and Biomolecular Engineering, Sogang University, 35 Baekbeom-Ro, Mapo-Gu, Seoul, 121-742, Republic of Korea;

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  • 正文语种 eng
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  • 关键词

    Enzyme-Ball; ELISA; Bovine Serum Albumin (BSA); Horseradish Peroxidase (HRP);

    机译：酶球ELISA;牛血清白蛋白（BSA）;辣根过氧化物酶（HRP）;

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