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Development of a novel nanomaterial-based optical platform for a protease biosensor

机译:新型基于纳米材料的蛋白酶生物传感器光学平台的开发

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We have been investigating a new nanomaterial-based optical platform for the immobilization of protease substrates for the development of a biosensor to detect medically relevant enzymes. Metallic nanoparticles have been deposited onto thin films and are being used for their optical properties. Two different peptide sequences have been designed as trypsin substrates that are designed to be immobilized onto the surface of the thin films. The peptides were synthesized with a fluorophore attached at the terminal end of the peptide to allow for fluorescence sensing. Fluorescent molecules in close proximity to metallic elements will have their fluorescence signal quenched due to surface plasmon resonance (SPR) effects. When the peptide is cleaved by trypsin, the fluorophore is separated, resulting in a detectable change in fluorescence intensity. These novel nanomaterial-based optical platforms have been fabricated using physical vapor deposition. Innovative techniques have been invented using these machines to acquire nanoparticles in the range of a few nanometers on these thin films. It is known that nanoparticles with dimensions less than their bandwidth display optical properties much different from their bulk counterparts. We have immobilized the peptide substrates to the surface of the metallized thin films so they are in close proximity with each other. Polydimethlysiloxane (PDMS) was molded to create small wells and placed on the thin films. Fluorescent microscopy was used to image the wells as various concentrations of the enzyme were introduced resulting in a recovery of green fluorescence from the fluorophore on the cleaved portion of the peptide. Different size nanoparticles and different immobilization processes are being used to optimize the design of the protease biosensor.
机译:我们一直在研究一种新的基于纳米材料的光学平台,用于固定蛋白酶底物,以开发用于检测医学上相关酶的生物传感器。金属纳米颗粒已经沉积在薄膜上,并被用于其光学性能。已经设计了两种不同的肽序列作为胰蛋白酶底物,其被设计为固定在薄膜表面上。合成的肽在肽的末端带有一个荧光团,可以进行荧光传感。由于表面等离振子共振(SPR)效应,紧邻金属元素的荧光分子的荧光信号将被淬灭。当肽被胰蛋白酶切割时,荧光团被分离,导致荧光强度的可检测变化。这些新型的基于纳米材料的光学平台已使用物理气相沉积法制造。使用这些机器发明了创新技术,以在这些薄膜上获取几纳米范围内的纳米颗粒。众所周知,尺寸小于其带宽的纳米颗粒显示出的光学特性与其整体对应物有很大不同。我们已将肽底物固定在金属化薄膜的表面,因此它们彼此非常接近。将聚二甲基硅氧烷(PDMS)模制成小孔,然后放置在薄膜上。当引入各种浓度的酶时,使用荧光显微镜对孔进行成像,从而从肽的切割部分上的荧光团中回收绿色荧光。不同大小的纳米粒子和不同的固定过程被用于优化蛋白酶生物传感器的设计。

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