Peptide Diversity Generation Apparatus to Elucidate Cell-Molecules Interactions

研究细胞与其环境之间的相互作用需要分子工具的可用性来探测细胞环境并了解由此产生的细胞反应。为此，该提案的三位共同申请人利用药物化学（Eric Marsault），干细胞生物学（Florian Bentzinger）和生物物理学（Michel Grandbois）的互补专业知识来询问细胞。Marsault生产设计的细胞渗透剂，称为富含胍的转运蛋白，将其他不可渗透的货物输送到细胞，以调节细胞反应。重要的是，这些GRT必须是多样的，以探测细胞表面并找到特定的细胞识别元件。Bentzinger破译了肌肉干细胞及其环境的相互作用，以更好地了解干细胞生态位如何调节这些细胞的命运。最后，Grandbois开发了基于原子力显微镜和表面等离子体共振的方法和仪器，以询问单个细胞对异质细胞群体中不同刺激的反应。为了支持他们各自的NSERC资助的研究项目，共同申请人需要访问基础设施，以大量和微量生成多样化的肽和肽类似物阵列。与设计用于制备毫克或克规模的少量（几十个）肽的经典肽合成器相反，需要的是微克数量的大量（数百至数千个）肽。这是本申请的目的，即安装与载玻片点样机器人耦合的阵列肽合成器。目前缺乏这种设备严重限制了共同申请人研究计划的进展。所要求的设备不仅将解锁他们的项目并充分支持共同申请人的研究项目，还将作为与学术合作者和私营公司开始新合作的垫脚石。最后，它还将提供一个工具，为总部质量保证人员提供尖端技术，以增加他们的技能。

The study of interactions between cells and their environment requires the availability of molecular tools to probe the cellular environment and understand the resulting cellular response. Toward this end, the three co-applicants on this proposal use complementary expertise in medicinal chemistry (Eric Marsault), stem cell biology (Florian Bentzinger) and biophysics (Michel Grandbois) to interrogate cells.***Marsault generates designer cell penetration agents known as guanidine-rich transporters to deliver otherwise impermeable cargos to cells to modulate cellular response. Importantly, these GRTs have to be diverse to probe the cellular surface and find specific cellular recognition elements.***Bentzinger deciphers the interactions of muscle stem cells and their environment to better understand how stem cells niche modulates the fate of these cells. Finally, Grandbois develops atomic force microscopy and surface plasmon resonance-based methodologies and instruments to interrogate the response of individual cells to different stimuli in heterogenous cell populations.******In order to support their individual NSERC-funded research programs, the co-applicants require access to an infrastructure to generate diversified arrays of peptides and peptidic analogs, in large numbers and micro-quantities. In contrast to a classical peptide synthesizer designed to prepare small numbers (dozens) of peptides in milligram or gram scales, the needs are for large numbers (hundreds to thousands) of peptides in microgram quantities. This is the purpose of this application, i.e. to install an array peptide synthesizer coupled to a slide spotting robot. The current lack of this equipment severely restricts advances in the research programs of the co-applicants. The requested equipment will not only unlock their programs and adequately support the research program of the co-applicants, it will also serve as a stepping stone to start new collaborations with academic collaborators and private companies. Finally, it will also provide an instrument that will provide HQPs with cutting edge technology to be added to their skillset.