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

这个PFI：AIR技术翻译项目专注于翻译蛋白质组学领域的新技术的开发。蛋白质组学是对生物、组织或细胞产生的蛋白质进行高通量研究。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：哪些蛋白质在正常发育和疾病状态之间存在差异，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的生物标志物。蛋白质组学方法已经在大量的条件下应用于广泛的生物体。这是一个相对较新的领域，要充分发挥其潜力，需要跨越许多技术障碍。在蛋白质组学领域工作的科学家们表示，蛋白质和多肽样品的制备是他们面临的最大实验障碍之一。在美国，每年有100多万个这样的蛋白质组样本准备进行分析，可重复性、可靠性和自动化的样本准备非常重要。目前，现有的样品制备方法缺乏普适性，偏向于特定类别的蛋白质和多肽，导致大量样品损失，而且现有方法难以实现工业应用的自动化。该奖项开发了“通用蛋白质和多肽分离和清理试剂盒”(UPICK)的原型，其目标是所有蛋白质和多肽的普遍特征，因此它是通用和公正的。UPICK易于实现自动化，使其能够应用于高吞吐量的工业工作流程，同时将处理错误和环境污染降至最低。UPICK将允许研究人员以更高的可靠性、可重复性和标准化来执行他们的蛋白质组实验，从而加强生物医学发现。该项目解决了以下技术差距(S)，因为它从研究发现转化为商业应用。将为蛋白质清除试剂盒开发两种新的试剂。一个是开发一种完全可逆的蛋白质/肽标签，它能够在化学上偶联到第二个新特征，即超高容量磁珠。这些试剂将首先用于标记溶液中的所有蛋白质或多肽，然后将标记的蛋白质/多肽共价结合到磁珠上。然后对珠子进行清洗，以去除内源性和外源性的非蛋白质污染物，如DNA、RNA、盐、洗涤剂和变性剂。目标是创建一个UPICK原型来分发以进行Beta测试。研究生和本科生团队将参与开发试剂盒，并在内部和与Beta测试伙伴一起测试试剂盒的有效性和易用性。此外，学生还将有机会参加卡内基梅隆大学S商学院商业创新学者项目，该项目旨在培养非商学院研究生的创业技能。

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.