Multiplexed Tracking of Nanoparticles using Laser Desorption/Ionization Mass Spec

描述（由申请人提供）：纳米颗粒与生物系统的相互作用是生物医学和环境科学中的一个重要问题。在我们提出的研究中，我们将利用选择性激光解吸和离子化的配体连接到纳米材料，以跟踪细胞摄取和内体释放的纳米材料使用质谱。在这种方法中，配体充当质量“条形码”，允许同时跟踪多个颗粒。 在我们提出的研究中，我们将采取多管齐下的方法来系统研究细胞对纳米颗粒的吸收，将分析方法学的发展与合成和生物学相结合。我们的研究具有三个目标，将并行追求：目标1：Rotello将制造纳米粒子的大小和表面功能的系统变化。目标二：Rotello将以多重方式与不同细胞类型、相同细胞类型的不同细胞系以及特定细胞系的同基因变体孵育颗粒。然后，Vachet将使用LDI来确定相对和绝对颗粒摄取。一旦确定了细胞摄取的基本参数，Rotello和Vachet将使用LDI-MS来定量内体释放。这些研究将为纳米材料的细胞摄取和内体释放提供定量结构-活性相关性。目标3：Vachet将参数化地探索优化LDI颗粒跟踪灵敏度的方法，并将研究该方法的分析范围。这些结果将提供增强的探针，为未来的应用LDI在体内的粒子跟踪。我们的研究将把Rotello小组开发的纳米颗粒合成和生物学能力与Vachet的仪器和定量能力结合起来。这项R21资助的目标是双重的：开发和验证纳米颗粒细胞摄取的多重分析，并应用该方法探索纳米材料细胞摄取的结构-活性特性。这些结果本身在交付和诊断应用方面将是重要的。此外，他们将奠定基础，应用激光多普勒成像的纳米粒子在体内和环境中的跟踪。 公共卫生相关性：跟踪纳米粒子是生物医学和环境科学中的一个重要问题。我们将开发一种策略，允许同时跟踪多种类型的纳米颗粒，并使用这种方法来表征这些材料的细胞摄取。

DESCRIPTION (provided by applicant): The interaction of nanoparticles with biological systems is an important issue in biomedical and environmental sciences. In our proposed research we will exploit the selective laser desorption and ionization of ligands attached to nanomaterials to track cellular uptake and endosomal release of nanomaterials using mass spectrometry. In this approach the ligands serve as mass "barcodes", allowing the simultaneous tracking of multiple particles. In our proposed research we will take a multi-pronged approach to the systematic study of cellular uptake of nanoparticles that integrates analytical methodology development with synthesis and biology. Our research features three Aims that will be pursued in parallel: Aim 1: Rotello will fabricate nanoparticle with systematic variations in size and surface functionality. Aim 2: Rotello will incubate particles in multiplex fashion with different cell types, different lines of the same cell type, and isogenic variants of specific cell lines. Vachet will then use LDI to determine relative and absolute particle uptake. Once the fundamental parameters for cellular uptake have been established, Rotello and Vachet will use LDI-MS to quantify endosomal release. These studies will provide quantitative structure-activity correlations for cellular uptake and endosomal release of nanomaterials. Aim 3: Vachet will parametrically explore methods to optimize the sensitivity of LDI particle tracking and will investigate the analytical scope of this method. These results will provide enhanced probes for future applications of LDI for particle tracking in vivo. Our research will couple the nanoparticle synthesis and biological proficiency developed by the Rotello group with the instrumental and quantitative capabilities of Vachet. The goal of this R21 grant is two-fold: development and validation of multiplex analysis of cellular uptake of nanoparticles coupled with the application of this method to explore structure-activity properties of cellular uptake of nanomaterials. These results will be important in and of themselves in terms of delivery and diagnostic applications. Moreover, they will lay the foundation for the application of LDI for the tracking of nanoparticles in vivo and in the environment. PUBLIC HEALTH RELEVANCE: Tracking nanoparticles is an important issue in biomedical and environmental science. We will develop a strategy that allows simultaneous tracking of multiple types of nanoparticles and use this method to characterize cellular uptake of these materials.