Innovating Bioanalytical Technologies using Bioinspired Nanomaterials

利用生物启发的纳米材料蛋白质创新生物分析技术是许多重要生物过程的核心，如调节细胞途径、催化代谢反应和肿瘤发生。量化细胞和生物体液中的蛋白质水平是表征蛋白质功能和调节生物过程的关键。拟议的研究计划将使用生物灵感纳米材料创新蛋白质分析技术，以追求我们解决生物分析化学中基本和应用挑战的愿景。这项研究计划本质上是跨学科的，将支持由分析化学家、材料化学家、临床化学家、工程师、生物学家和医学科学家组成的多样化团队。这项NSERC发现助学金计划将支持七名研究生，我们将通过三个目标实现我们的计划目标：目标1：确定生物和生物启发材料形成途径的基本原则。我们的目标是利用材料形成-结构-功能原理，战略性地设计用于蛋白质分析的生物启发纳米材料。我们假设，纳米尺度的相互作用和相决定了纳米材料的形成路径，并指导了纳米材料的结构和功能。我们建议使用一种新的互补方法来研究生物矿物和生物启发纳米材料的形成和功能，该方法结合了单分子荧光和低温电子显微镜。在短期内，我们将研究酶基磷酸钙和催化活性金属有机骨架(MOF)的形成机理。目的2：展示用于蛋白质分析的概念验证互补的单分子分析方法。许多参与重要生物过程的蛋白质以非常低的浓度存在，需要足够灵敏的方法来检测少量分子。我的实验室将开发新的单分子技术，可以实现对多种蛋白质性质的超灵敏量化。在短期内，我们将开发两种概念验证分析方法，用于基于(1)金属纳米颗粒和(2)MOF纳米酶的酶浓度和活性的互补分析。目的：研制基于仿生纳米材料的复合蛋白质电化学生物传感器。实时蛋白质测量可以提供关于生物系统生理状态的动态信息，并可以通过具有快速响应时间的定量生物传感器获得。在短期内，我的实验室将开发基于MOF-抗体复合材料的电化学生物传感器，用于快速检测生物液中高度丰富的蛋白质。我们的目标是实现简单的工作流程、高灵敏度、快速响应时间和多路蛋白质测量。

Innovating Bioanalytical Technologies using Bioinspired Nanomaterials Proteins are central to many important biological processes such as regulating cellular pathways, catalyzing metabolic reactions, and tumorigenesis. Quantifying protein levels in cells and biofluids is critical to characterizing protein function and regulation of biological processes. The proposed research program will innovate protein-assay technologies using bioinspired nanomaterials to pursue our vision of solving fundamental and applied challenges in bioanalytical chemistry. This research program is inherently interdisciplinary and will support a diverse team of analytical chemists, material chemists, clinical chemists, engineers, biologist, and medical scientists. This NSERC Discovery Grant proposal will support seven graduate students and we will achieve our program goals through three objectives: Objective 1: Identify fundamental principles of biological and bioinspired material formation pathways. We aim to strategically design bioinspired nanomaterials for protein assays using material formation-structure-function principles. We hypothesize that nanoscale interactions and phases dictate formation pathways and direct the nanomaterial structure and function. We propose to study the formation and function of biomineral and bioinspired nanomaterials using a new complementary approach that integrates single-molecule fluorescence and cryotransmission electron microscopy. In the short term, we will investigate the formation mechanisms of enzyme-based calcium phosphate and of catalytically-active metal-organic frameworks (MOFs). Objective 2: Demonstrate proof-of-concept complementary, single-molecule assays for protein analyses. Many proteins that are involved in important biological processes exist at very low concentrations and require methodologies sensitive enough to detect a low number of molecules. My lab will develop new single-molecule technologies that can achieve ultrasensitive quantification of more than one protein property. In the short term, we will develop two proof-of-concept assays for the complementary analysis of enzyme concentration and activity based on (1) metal nanoparticles and (2) MOF nanozymes. Objective 3: Develop electrochemical biosensors based on bioinspired nanomaterials for multiplexed protein detection. Real-time protein measurements can provide dynamic information on the physiological state of a biological system and can be obtained via quantitative biosensors with rapid response times. In the short term, my lab will develop electrochemical biosensors based on MOF-antibody composites for rapid detection of highly-abundant proteins in biofluids. We aim to achieve simple workflows, high sensitivity, rapid response times, and multiplexed protein measurements.