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Novel ultra-stable enzymes and methods for proteomics

新型超稳定酶和蛋白质组学方法

基本信息

批准号：
10010437
负责人：
JILL O FUSS
金额：
$ 92.29万
依托单位：
CINDER BIOLOGICAL, INC.
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-01 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10010437
关键词：
Acids Address Archaea Biodiversity Biological Biological Process Businesses Characteristics Clinical Clinical Research Cloning Development Digestion Drops Effectiveness Enzymes Foundations Funding Funding Agency Future Gel Genetic Engineering Goals Health Hour Immobilization Laboratories Legal patent Liquid substance Maps Mass Spectrum Analysis Medical Medical Research Membrane Methods Microbe Modeling Modernization Peptide Hydrolases Performance Phase Planet Earth Preparation Prevention Procedures Process Production Protein Analysis Proteins Proteomics Protocols documentation Reaction Reaction Time Refractory Reproducibility Research Sales Sampling Science Services Specificity System Technology Testing Time Tissue Sample Traction Trypsin Urea Viral Water Work base biological research commercialization disease diagnosis empowered improved new technology novel phase 1 study product development recruit success tool

项目摘要

PROJECT SUMMARY/ABSTRACT The highly technical and rapidly advancing field of modern proteomics relies heavily on enzyme technologies that were established over one hundred and fifty years ago, namely trypsin. We propose to apply our entirely novel ultra-stable Archaeal proteases to make superior front-end digestion products for proteomics. Our patented technologies allow efficient production of ultra-stable enzymes with optima near the boiling point of water at highly acidic pH with half-lives on the order of weeks, not hours. The hot-acid conditions of CinderBio enzymatic digest reactions denatures target proteins and allows faster and more effective target digestion without added chaotropes like urea. Our hyperthermoacidic enzymes generate protein coverage and protein identification metrics that are competitive with overnight trypsin/urea digests with less sample, fewer steps, and five-minute reaction times. Here we propose to build on over a decade of our work with these enzyme systems to commercialize proteomic digestion products with novel capabilities and cleavage specificities to empower proteomic research. We have established academic and corporate partnerships to facilitate this work to expedite product development and deployment of our novel capabilities into the proteomics markets within the next two years. With the astonishing rate of technical advances in proteomics it is remarkable that front-end enzymatic digests have changed very little since the beginning of the field. Here we propose to bring to market novel enzyme products and capabilities to advance proteomic capabilities, reproducibility, and throughput with our efficient proteomic digestion products.

项目概要/摘要现代蛋白质组学这个技术含量高、发展迅速的领域在很大程度上依赖于酶一百五十多年前建立的技术，即胰蛋白酶。我们建议应用我们全新的超稳定古菌蛋白酶来实现卓越的前端消化蛋白质组学产品。我们的专利技术可以高效生产超稳定的酶在高酸性 pH 条件下，最佳值接近水的沸点，半衰期约为数周，而不是小时。 CinderBio 酶消化反应的热酸条件会使目标蛋白质变性，无需添加尿素等离液剂，即可实现更快、更有效的目标消化。我们的高温酸性酶产生蛋白质覆盖率和蛋白质识别指标与隔夜胰蛋白酶/尿素消化相比，样品更少，步骤更少，反应时间为五分钟次。在这里，我们建议以我们十多年来对这些酶系统的研究为基础将具有新颖功能和切割特异性的蛋白质组消化产品商业化增强蛋白质组学研究的能力。我们建立了学术和企业合作伙伴关系，以促进这项工作旨在加快产品开发并将我们的新颖功能部署到蛋白质组学中未来两年内的市场。随着蛋白质组学技术进步的惊人速度，值得注意的是，自该领域诞生以来，前端酶消化变化很小。在这里，我们建议将新颖的酶产品和能力推向市场，以推进蛋白质组学我们高效的蛋白质组消化产品的能力、重现性和通量。