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近红外荧光胰蛋白酶稳定金纳米簇作为表面等离子体增强能量转移生物传感器及体内癌症成像生物探针

Near infrared fluorescent trypsin stabilized gold nanoclusters as surface plasmon enhanced energy transfer biosensor and in vivo cancer imaging bioprobe.

机构信息

State Key Laboratory of Medicinal Chemical Biology, and Research Center for Analytical Sciences, College of Chemistry, Nankai University, Tianjin 300071, China.

出版信息

Anal Chem. 2013 Mar 19;85(6):3238-45. doi: 10.1021/ac303603f. Epub 2013 Mar 4.

Abstract

The simplicity of the green-synthesized routine and the availability of surface modification of diverse bioactive molecules make noble metal nanostructures highly suitable as multifunctional biomaterials for biological and biomedical application. Here, we report the preparation of trypsin stabilized gold nanoclusters (try-AuNCs) with near-infrared fluorescence for biosensing heparin based on surface plasmon enhanced energy transfer (SPEET) and folic acid (FA) modified try-AuNCs for in vivo cancer bioimaging. The SPEET/try-AuNCs fluorescence biosensor was designed via heparin mediated energy transfer between try-AuNCs and cysteamine modified gold nanoparticles (cyst-AuNPs). The developed SPEET/try-AuNCs fluorescence biosensor allowed sensitive and selective detection of heparin with a linear range of 0.1-4.0 μg mL(-1) and a detection limit (3s) of 0.05 μg mL(-1). The relative standard deviation for eleven replicate detections of 2.5 μg mL(-1) heparin was 1.1%, and the recoveries of the spiked heparin in human serum samples ranged from 97% to 100%. In addition, folic acid was immobilized on the surface of try-AuNCs to ameliorate the specific affinity of AuNCs for tumors, and the near-infrared fluorescent FA-try-AuNCs were applied for in vivo cancer imaging of high folate receptor (FR) expressing Hela tumor. In vivo study of the dynamic behavior and targeting ability of FA-try-AuNCs probe to Hela tumor bearing mice and normal nude mice validated the high specific affinity of FA-try-AuNCs probe to FR positive tumors. The results show that the prepared try-AuNCs have great potential as multifunctional biomaterials for biosensing biomolecules with SPEET mode and in vivo cancer imaging with high targeting ability.

摘要

绿色合成方法的简单性以及多种生物活性分子的表面修饰可用性,使得贵金属纳米结构非常适合作为多功能生物材料,用于生物和生物医学应用。在这里,我们报告了一种基于表面等离子体增强能量转移(SPEET)的近红外荧光生物传感肝素的胰蛋白酶稳定金纳米簇(try-AuNCs)的制备方法,以及叶酸(FA)修饰的 try-AuNCs 用于体内癌症生物成像。通过肝素介导的 try-AuNCs 和半胱氨酸修饰的金纳米粒子(cyst-AuNPs)之间的能量转移,设计了 SPEET/try-AuNCs 荧光生物传感器。开发的 SPEET/try-AuNCs 荧光生物传感器允许对肝素进行灵敏和选择性检测,线性范围为 0.1-4.0 μg mL(-1),检测限(3s)为 0.05 μg mL(-1)。2.5 μg mL(-1)肝素的 11 个重复检测的相对标准偏差为 1.1%,人血清样品中肝素的加标回收率在 97%-100%之间。此外,叶酸被固定在 try-AuNCs 的表面上,以改善 AuNCs 对肿瘤的特异性亲和力,并且近红外荧光 FA-try-AuNCs 被应用于高叶酸受体(FR)表达的 Hela 肿瘤的体内癌症成像。FA-try-AuNCs 探针在荷瘤 Hela 小鼠和正常裸鼠体内的动态行为和靶向能力的体内研究验证了 FA-try-AuNCs 探针对 FR 阳性肿瘤的高特异性亲和力。结果表明,所制备的 try-AuNCs 具有作为多功能生物材料的巨大潜力,可用于基于 SPEET 模式的生物传感生物分子和具有高靶向能力的体内癌症成像。

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