Inorganic Chemistry Tools for Bioconjugation, Recognition and Imaging

用于生物共轭、识别和成像的无机化学工具

• 批准号: 10406790
• 负责人: Alexander Michael Spokoyny
• 金额: $ 37.09万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10406790
• 关键词: 3-Dimensional; Address; Affinity; Area; Binding; Biological; Boron; Cells; Chemistry; Communities; Development; Diagnostic; Disease; Electron Microscopy; Fluorescent Dyes; Funding; Gold; Growth Factor; Hybrids; Image; Imaging Techniques; Inorganic Chemistry; Kinetics; Knowledge; Label; Laboratories; Lead; Ligands; Membrane; Metals; Methods; Modernization; Modification; Molecular; Organelles; Peptides; Positioning Attribute; Positron-Emission Tomography; Process; Protein Inhibition; Proteins; Reaction; Reagent; Research; Stains; Sulfhydryl Compounds; Surface; Therapeutic; Virus; Work; base; biomaterial compatibility; cell growth; combat; covalent bond; fluorophore; functional group; interest; light microscopy; metallicity; microscopic imaging; multimodality; nanoGold; nanoparticle; receptor; scaffold; small molecule; tool

Project Summary/Abstract Our laboratory is interested in developing new inorganic chemistry tools to address unmet needs in the areas of bioconjugation, recognition, and imaging. In order to tackle these challenges, new molecular scaffolds and biocompatible chemistry are crucial. The overall objective of this competitive MIRA renewal application is to further advance the field of organomimetic boron cluster chemistry. Organomimetic features of these clusters arise from 1) their ability to undergo facile functionalization chemistry with a wide array of substituents, forming stable covalent bonds attached to the cluster¢s vertices and 2) unique 3D aromaticity, rendering these clusters amenable for several modes of microscopy imaging. Within the scope of this work is also a set of new, rapid organometallic transformations that were discovered during the original cycle of the MIRA funding that would allow one to rationally tether boron clusters and other molecules under biologically relevant conditions. Our laboratory is interested in developing new transformations that mimic the operational simplicity with which thiol ligands normally assemble onto a metallic gold surface. This chemistry has previously revolutionized the ease with which we can create hybrid noble metal nanoparticles (e.g., thiol capped gold nanoparticles - AuNPs). However, these hybrid AuNPs are not atomically precise, and the ligand corona is dynamic. These features lead to hybrids with a non-uniform composition and size, ultimately limiting their applications for the inhibition of protein-biomolecule interactions. Addressing this challenge, we have developed organometallic- based methods for cluster modification, providing a covalently tethered dense corona of functional biomolecules and ligands spatially arranged with three-dimensional precision. We propose to further expand this approach to rapidly build up sophisticated atomically-precise 3D nanomolecules for multivalent binding to various biological targets, including virus entry receptors, biological membranes, and cellular growth factors. We also propose to utilize the inherent robustness and reaction kinetics associated with the developed Au-based reagents for biomolecular positron-emission tomography (PET) labeling. We have also been engaged in the development of new boron cluster chemistry, allowing for the positioning of multiple reactive functional groups on a three- dimensional cluster and use these rigid 3D species to label and tether biomolecules to achieve unconventional folding and recognition. We propose chemistry that will enable the labeling of small molecules, peptides, proteins, and cells with various boron cluster scaffolds, which can subsequently be used as multivalent binders, affinity tags, and fluorescent-free labels using Raman microscopy imaging. Lastly, we propose the use of perfunctionalized boron clusters as dual staining/fluorophore agents for correlative light and electron microscopy (CLEM). Specifically, we will work on developing hybrids that contain heavy atom perfunctionalized boron cluster stains tethered to a fluorescent dye, which would enable targeting of specific cellular organelles, thereby providing access to an unprecedented class of photostable and non-toxic agents for CLEM.

项目总结/摘要 我们的实验室有兴趣开发新的无机化学工具，以解决未满足的需求， 生物结合、识别和成像领域。为了应对这些挑战，新的分子支架 和生物相容性化学是至关重要的这种竞争MIRA续期申请的总体目标是 进一步推进有机模拟硼原子簇化学领域。这些簇的有机模拟特征 1）它们能够用广泛的取代基进行容易的官能化化学， 稳定的共价键连接到簇的顶点和2）独特的三维芳香性，使这些簇 适合于几种显微镜成像模式。在这项工作的范围内也是一套新的，快速的 在MIRA资助的原始周期中发现的有机金属转化， 允许人们在生物学相关条件下合理地束缚硼簇和其他分子。 我们的实验室有兴趣开发新的转换，模仿操作简单性， 所述硫醇配体通常组装到金属金表面上。这种化学反应在以前已经彻底改变了 我们可以容易地产生混合贵金属纳米颗粒（例如，硫醇封端的金纳米颗粒- AuNP）。然而，这些混合金纳米粒子不是原子精确的，并且配体电晕是动态的。这些 这些特征导致混合物具有不均匀的组成和尺寸，最终限制了它们在生物医学领域的应用。 抑制蛋白质-生物分子相互作用。为了应对这一挑战，我们开发了有机金属- 基于簇修饰的方法，提供功能性生物分子的共价束缚的致密冠 和以三维精度空间排列的配体。我们建议进一步扩大这一做法， 快速构建复杂的原子级精确的3D纳米分子，用于多价结合各种生物学 靶点，包括病毒进入受体、生物膜和细胞生长因子。我们亦建议 利用与开发的Au基试剂相关的固有鲁棒性和反应动力学， 生物分子正电子发射断层扫描（PET）标记。我们还参与了 新的硼簇化学，允许在三个- 三维集群，并使用这些刚性的3D物种来标记和拴系生物分子，以实现非常规的 折叠和识别。我们提出了一种化学方法，可以标记小分子，肽，蛋白质， 以及具有各种硼簇支架的细胞，其随后可用作多价结合剂， 标签和无荧光标记，使用拉曼显微成像。最后，我们建议使用 用于相关光学和电子显微术的作为双染色/荧光团试剂的全官能化硼簇 （CLEM）。具体来说，我们将致力于开发包含重原子全官能化硼簇的杂化物 染料拴在荧光染料上，这将使特定的细胞器成为目标， 为CLEM提供了前所未有的光稳定和无毒试剂。