PFI:AIR - TT: Development of a universal protein and peptide cleanup kit

PFI:AIR - TT：开发通用蛋白质和肽清理试剂盒

• 批准号: 1700833
• 负责人: Jonathan Minden
• 金额: $ 20万
• 依托单位: Carnegie-Mellon University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-15 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1700833&HistoricalAwards=false
• 关键词: PFI; AIR; TT; Development; universal

This PFI: AIR Technology Translation project focuses on translating the development of novel technologies for the field of proteomics. Proteomics is the high throughput study of proteins produced by organisms, tissues or cells. Proteomics methods enable biomedical researchers in industry and academia to discover the following knowledge: what proteins differ between normal development and diseased states, what proteins are affected by exposure to different drugs or treatments, and what proteins potentially serve as disease biomarkers. Proteomics methods have been applied to a wide range of organisms under an enormous number of conditions. It a relatively new field that has numerous technological hurdles to cross in order to meet its full potential. Scientists working in the field of proteomics have indicated that protein and peptide sample preparation is one of the greatest experimental hurdles they face. More than a million of such proteomics samples are prepared for analysis per year in the U.S. and reproducible, reliable and automatable sample preparation is important. Currently, the existing sample preparation methods lack universality, they are biased towards particular classes of proteins and peptides that results in significant sample loss, and the current methods are difficult to automate for industrial applications. This award develops a prototype of a "Universal Protein and Peptide Isolation and Cleanup Kit" (UPICK) that targets a ubiquitous feature of all proteins and peptides, thus it is universal and unbiased. UPICK is amenable to automation, enabling adoption into high-throughput, industrial workflows, while minimizing handling errors and environmental contamination. UPICK will allow researchers to perform their proteomics experiments with far greater reliability, repeatability and standardization, thus enhancing biomedical discovery. This project addresses the following technology gap(s) as it translates from research discovery toward commercial application. Two novel reagents will be developed for the protein cleanup kit. One is the development of a completely reversible protein/peptide tag that is capable of chemically coupling to the second novel feature, which is ultra-high capacity magnetic beads. Together, these reagents will be used to first tag all proteins or peptides in solution and then covalently bind the tagged proteins/peptides to magnetic beads. The beads will then be washed to remove endogenous and exogenous non-protein contaminants, such as DNA, RNA, salts, detergents and denaturants. The goal is to create a UPICK prototype to distribute for beta-testing. Graduate students and a team of undergraduates will be involved in developing the kits and testing the efficacy and ease of use of the kits both in-house and with beta-test partners. In addition, the students will have the opportunity to participate in Carnegie Mellon?s Tepper School of Business Innovation Scholars Program that develops the entrepreneurial skills of non-business school, graduate students.

这个PFI：AIR技术翻译项目的重点是翻译蛋白质组学领域的新技术的发展。蛋白质组学是对生物体、组织或细胞产生的蛋白质的高通量研究。蛋白质组学方法使工业界和学术界的生物医学研究人员能够发现以下知识：正常发育和疾病状态之间有哪些蛋白质不同，哪些蛋白质受到不同药物或治疗的影响，以及哪些蛋白质可能作为疾病生物标志物。蛋白质组学方法已经在大量条件下应用于广泛的生物体。这是一个相对较新的领域，为了充分发挥其潜力，有许多技术障碍需要跨越。蛋白质组学领域的科学家已经指出，蛋白质和肽样品制备是他们面临的最大实验障碍之一。在美国，每年有超过100万个这样的蛋白质组学样品被制备用于分析，可重复、可靠和自动化的样品制备非常重要。目前，现有的样品制备方法缺乏通用性，它们偏向于特定类别的蛋白质和肽，导致显著的样品损失，并且当前的方法难以自动化用于工业应用。该奖项开发了一种“通用蛋白质和肽分离和纯化试剂盒”（UPICK）的原型，该试剂盒针对所有蛋白质和肽的普遍存在的特征，因此它是通用和无偏见的。UPICK可实现自动化，可用于高通量的工业工作流程，同时最大限度地减少处理错误和环境污染。UPICK将使研究人员能够以更高的可靠性，可重复性和标准化进行蛋白质组学实验，从而提高生物医学发现。该项目解决了以下技术差距，因为它从研究发现转化为商业应用。两种新型试剂将被开发用于蛋白质清除试剂盒。一个是开发一种完全可逆的蛋白质/肽标签，该标签能够与第二个新功能（超高容量磁珠）化学偶联。这些试剂将一起用于首先标记溶液中的所有蛋白质或肽，然后将标记的蛋白质/肽共价结合到磁珠上。然后洗涤珠子以去除内源性和外源性非蛋白质污染物，例如DNA、RNA、盐、洗涤剂和变性剂。我们的目标是创建一个UPICK原型，以进行beta测试。研究生和一个本科生团队将参与开发工具包，并在内部和与beta测试合作伙伴一起测试工具包的功效和易用性。此外，学生将有机会参加卡内基梅隆大学？Tepper商学院的创新学者项目旨在培养非商学院研究生的创业技能。