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Multiplexed proteomics-based kinase assay development

基于多重蛋白质组学的激酶测定开发

基本信息

批准号：
10793244
负责人：
Laurie L. Parker
金额：
$ 11.3万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2026-04-30
项目状态：
未结题

项目摘要

Project Summary This supplement application is to request a MultiPep2 peptide library synthesizer from CEM Corporation, to be used for preparing libraries of synthetic peptide candidates as a critical part of our efforts in optimal kinase substrate peptide design. The major, overarching goal of this R01 project is to develop panels of cell-deliverable peptide substrates for kinases that will enable identification of signatures of signaling activity in live cells using high-throughput targeted mass spectrometry. There are two components of this development in progress: · Kinase substrate preference mapping, and peptide substrate design based on those preferences · Focused selection of substrates that perform well in mass spectrometry detection Both of these goals require screening of many peptides to determine the best candidates to define a given kinase's activity, which we will eventually establish as assays using the targeted mass spectrometry approaches that are in development with our collaborator at Cedars Sinai. The MultiPep2 instrument is a library-scale peptide synthesizer, with versatile capabilities to synthesize up to 384 peptides per run, at a small multiwell- plate scale that is far more resource and reagent-efficient for this purpose than our current production-scale, non-library synthesizer, which can only make up to 24 at a time at about 10X greater scale and thus reagent and solvent usage. This will enhance the project's efficiency in screening peptide candidates, reduce solvent usage, associated reagent costs, and environmental impact, and reduce researcher time on library synthesis, reserving it for more intellectually demanding aspects of the work.

项目概要本补充申请是向CEM公司索取MultiPep2肽库合成仪，用于制备合成肽候选物文库，作为我们优化激酶工作的关键部分底物肽设计。该 R01 项目的主要总体目标是开发可交付细胞的面板激酶的肽底物将能够使用以下方法识别活细胞中信号活动的特征高通量靶向质谱分析。该开发有两个部分正在进行中： · 激酶底物偏好图谱，以及基于这些偏好的肽底物设计 · 重点选择质谱检测性能良好的底物这两个目标都需要筛选许多肽，以确定定义给定特定肽的最佳候选肽。激酶的活性，我们最终将使用靶向质谱方法进行测定正在与我们在 Cedars Sinai 的合作者一起开发。 MultiPep2 仪器是图书馆规模的仪器肽合成仪，具有多种功能，每次运行可在小型多孔孔中合成多达 384 个肽与我们目前的生产规模相比，平板规模的资源和试剂效率更高，非文库合成器，一次最多只能合成 24 个，规模约为 10 倍，因此试剂和溶剂的使用。这将提高项目筛选候选肽的效率，减少溶剂使用量、相关试剂成本和环境影响，并减少研究人员进行文库合成的时间，将其保留用于工作中对智力要求更高的方面。