SBIR Phase II: Universal Proteome Sample Preparation Kit Development

点击翻译按钮获取中文摘要

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project is to develop a universal protein sample preparation kit that extracts proteins from any biological sample, removes contaminants, and prepares them for downstream analytical methods with higher yields and deeper coverage than currently available technology. Proteins are the molecular machines of the cell that do all the work required to keep humans healthy, and incorporation of protein analysis is becoming increasingly important for developing a clear understanding of integrated biological systems, realizing the promise of precision medicine, and rapid drug discovery. Sample preparation is the first step in any protein-based project workflow and without a high-yield, fast, and reproducible way to prepare samples, downstream analyses are difficult to interpret and highly variable, leading to long project times and wasted resources. The proposed innovation improves sample yields, reproducibility, and speed, accelerating research seeking to discover new therapies and diagnostics and improving cost-effectiveness. It will also purify other important biomolecules from the same sample, such as DNA, RNA, and metabolites, allowing researchers to obtain more information from each sample, prepare multiple samples in the time it takes to prepare one, and improve reproducibility by using the same starting material. The proposed project will utilize reversible protein tagging chemistry to create a one-tube workflow for the capture, wash, and elution of protein samples for analysis. Because this novel reversible protein tagging chemistry is not dependent on a catalyst, does not produce byproducts, and is completely biorthogonal, it has the potential to work universally with all types of buffer systems, including harsh denaturants and detergents commonly used for cell lysis and protein solubilization. This proposed work will optimize the utilization of this sample preparation technology in the most used buffer systems, model organisms, and will show superior yields in the preparation of diagnostic sample types such as blood, urine, and cerebrospinal fluid. Additionally, because our reversible protein tag does not cross react with other biomolecules, we will show that we can also use this technology to purify other important biomolecules such as DNA, RNA, and metabolites. The technology developed in this project will revolutionize how researchers prepare samples for analysis, providing them with modular tools that can purify any biomolecules they need from the same starting sample, reducing time, improving yields, and decreasing batch to batch variability by comparing within the same starting sample.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.