Energy transfer-based multiplexed assay of proteases by using gold nanoparticle and quantum dot conjugates on a surface
Energy transfer-based multiplexed assay of proteases by using gold nanoparticle and quantum dot conjugates on a surface
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DOI：
10.1021/ac702416e
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发表时间：
2008-06-15
影响因子：
7.4
通讯作者：
Kim, Hak-Sung
中科院分区：
文献类型：
--
作者：
Kim, Young-Pil;Oh, Young-Hee;Kim, Hak-Sung
快速、灵敏、高通量地检测蛋白酶及其抑制作用在诊断和制药领域具有重要意义。我们开发了一种多重检测系统的蛋白酶和它们的抑制，通过测量量子点（QD）和金纳米粒子（AuNPs）之间的能量转移在载玻片上。在该系统中，虽然固定在表面上的供体QD的光致发光（PL）由于紧邻的AuNP（能量受体）的存在而被淬灭，但蛋白酶活性引起受体和供体之间的能量转移效率的调节，从而使得能够进行蛋白酶测定。与量子点-染料体系相比，量子点-金纳米粒子的偶联物产生了更高的能量转移效率，导致蛋白酶的定量测定具有更高的灵敏度。当测试基质金属蛋白酶、半胱天冬酶和凝血酶时，成功地实现了多重测定，因为AuNP可以用作与具有不同颜色的QD结合的共同能量受体。我们的系统有望在蛋白酶相关疾病的诊断和高通量高灵敏度的潜在药物筛选中找到应用。
Rapid and sensitive assay of proteases and their inhibition in a high-throughput manner is of great significance in the diagnostic and pharmaceutical fields. We developed a multiplexed assay system of proteases and their inhibition by measuring the energy transfer between quantum dots (QDs) and gold nanoparticles (AuNPs) on a glass slide. In this system, while the photoluminescence (PL) of donor QDs immobilized on a surface was quenched due to the presence of AuNPs (energy acceptor) in close proximity, the protease activity caused modulation in the efficiency of the energy transfer between the acceptor and donor, thus enabling the protease assay. In comparison to the QD-dye system, the conjugate of the QD-AuNP gave rise to higher energy transfer efficiency, resulting in quantitative assay of proteases with more sensitivity. When matrix metalloproteinase, caspase, and thrombin were tested, a multiplexed assay was successfully achieved since the AuNP could be used as a common energy acceptor in conjunction with QDs having different colors. Our system is anticipated to find applications in the diagnosis of protease-related diseases and screening of potential drugs with high sensitivity in a high-throughput way.