Dextran-based polymeric chemiluminescent compounds for the sensitive optical imaging of a cytochrome p450 protein on a solid-phase membrane.
Dextran-based polymeric chemiluminescent compounds for the sensitive optical imaging of a cytochrome p450 protein on a solid-phase membrane.
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DOI：
10.1002/anie.200702290
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发表时间：
2007-11
影响因子：
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通讯作者：
Huan Zhang;Chaivat Smanmoo;T. Kabashima;Jianzhong Lu;M. Kai
中科院分区：
文献类型：
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作者：
Huan Zhang;Chaivat Smanmoo;T. Kabashima;Jianzhong Lu;M. Kai
化学发光 (CL) 已在许多科学领域得到广泛应用。[1] CL 成像是一种很有前景的检测系统，越来越多地用于多种分析物的超灵敏定量和定位。目前，微阵列技术因其在同时检测多种分析物方面的优势而在生物样品分析中得到普及。 [2] CL 信号通常由电荷耦合器件 (CCD) 相机测量，然后由计算机中的成像软件量化。对于化学发光成像，传统方法通常采用辣根过氧化物酶（HRP）或碱性磷酸酶作为信号酶，尽管该酶在室温下稳定性降低和高背景干扰限制了该技术在临床分析中的适用性，特别是对于血清样本。 [3]因此，鼓励开发非酶化学发光成像探针。在此，我们报告了一种合成基于葡聚糖的化学发光化合物的简单方法，及其作为化学发光标记大分子探针的应用，用于聚偏二氟乙烯（PVDF）膜上细胞色素P450（CYP）蛋白的灵敏化学发光成像（图1）。基于葡聚糖的化学发光化合物与少量生物素作为连接体和大量鲁米诺或异鲁米诺作为CL发射体连接。鲁米诺和异鲁米诺因其 CL 特性而闻名，并且其机理细节已被描述。 [4]此外，亲和素-生物素相互作用已在免疫组织化学、酶联免疫吸附测定和分子生物学中得到认可。 [5]生物素与亲和素结合的亲和力极高，缔合常数为 1015 m-1。为了获得良好的信号强度和高灵敏度，需要对鲁米诺和异鲁米诺衍生物的合成进行广泛的研究，以寻找一种新型的非酶探针。我们首先合成了含鲁米诺或异鲁米诺
Chemiluminescence (CL) has been exploited within a wide range of applications in many scientific fields.[1] CL imaging represents a promising detection system that is increasingly used for the ultrasensitive quantification and localization of several analytes. Currently, microarray technology has gained in popularity for the analysis of biological samples because of its benefits in the simultaneous detection of multiple analytes.[2] A CL signal is generally measured by a charge-coupled device (CCD) camera and then quantified by imaging software in a computer. For CL imaging, the traditional method usually employs horseradish peroxidase (HRP) or alkaline phosphatase as a signal enzyme, although the reduced stability of the enzyme at room temperature and high background interference limit the applicability of the technique in clinical analyses, especially for serum samples.[3] Thus, the development of a nonenzymatic CL-imaging probe is encouraged.Herein, we report a simple method for synthesizing dextran-based chemiluminescent compounds and their application as CL-labeling macromolecular probes for the sensitive CL imaging of a cytochromeP450 (CYP) protein on a poly (vinylidene difluoride)(PVDF) membrane (Figure1). The dextran-based chemiluminescent compound was tethered with a small amount of biotin as a linker and a large amount of luminol or isoluminol as CL emitter. Luminol and isoluminol are known for their CL properties, and their mechanistic details have been described.[4] In addition, the avidin–biotin interaction has been recognized in immunohistochemistry, enzyme-linked immunosorbent assay, and molecular biology.[5] The affinity of biotin binding to avidin is extremely high with an association constant of 1015 mÀ1. To obtain a good signal strength and high sensitivity, extensive work on the synthesis of luminol and isoluminol derivatives is desirable to search for a novel nonenzymatic probe. We first synthesized luminol-or isoluminol-containing