Microcapsule biosensors using competitive binding resonance energy transfer assays based on apoenzymes
Microcapsule biosensors using competitive binding resonance energy transfer assays based on apoenzymes
复制标题
DOI：
10.1021/ac050755u
复制
发表时间：
2005-09-01
影响因子：
7.4
通讯作者：
McShane, MJ
中科院分区：
文献类型：
--
作者：
Chinnayelka, S;McShane, MJ
本文报道了基于荧光共振能量转移的葡萄糖传感器的首次演示，其中竞争性结合（CB）测定被封装到微胶囊中。这项工作支持了这样的概念，即微胶囊是上级的水凝胶系统或其他基质的竞争性结合为基础的系统，因为它们提供了胶囊内部的传感元件的自由移动，同时保持恒定的总传感测定浓度。在这些初步实验中采用的转导方法也是一种新的CB系统的基础上的模型脱辅基酶，脱辅基葡萄糖氧化酶（AG），这是高度特异性的β-D-葡萄糖，作为模型的目标结合蛋白。包封在微胶囊中的异硫氰酸荧光素（FITC）-葡聚糖和异硫氰酸四甲基罗丹明-AG的葡萄糖敏感性显示出对β-D-葡萄糖的特异性比其他糖高5倍，中敏感地观察到，响应的灵敏度和范围可以通过使用不同的FITC标记物控制测定浓度来定制。葡聚糖分子量和总胶囊浓度。研究结果支持的概念，使用微胶囊封装CB检测可逆和稳定的传感器和使用脱辅基酶作为特定的分子识别元件在CB检测。此外，微胶囊葡萄糖传感器的表征结果证明了其适用于监测生理葡萄糖水平。
This paper reports the first demonstration of a fluorescence resonance energy transfer based glucose sensor, wherein a competitive binding (CB) assay is encapsulated into polyelectrolyte microcapsules. The work supports the concept that microcapsules are superior to hydrogel systems or other matrixes for competitive-binding-based system, as they provide free movement of the sensing elements within the capsule interior while constant total sensing assay concentration is maintained. The transduction approach employed in these preliminary experiments is also a novel CB system based on a model apoenzyme, apo-glucose oxidase (AG), which is highly specific to beta-D-glucose, as the model target-binding protein. The glucose sensitivity of the fluorescein isothiocyanate (FITC)-dextran and tetramethylrhodamine isothiocyanate-AG encapsulated in microcapsules showed 5 times greater specificity for beta-D-glucose over other sugars, with sensitivity (change in intensity ratio) in the range of 2-6%/mM. It was observed that the sensitivity and range of the response can be tailored by controlling the assay concentration using different FITC-dextran molecular weight and total capsule concentration. The findings support the concepts of using microcapsules to encapsulate CB assays for reversible and stable sensors and the use of apoenzymes as specific molecular recognition elements in CB assays. Further, characterization results for microcapsule glucose sensors demonstrate their suitability for monitoring physiological glucose levels.