PFI: TT: Development of 5-Minute Protein Analyses

该创新-技术转化伙伴关系（PFI-TT）项目的更广泛影响/商业潜力包括开发一种方便的设备，可以在5分钟内确定蛋白质浓度。 目前的蛋白质分析方法通常采用昂贵的试剂盒，需要超过1小时才能获得结果。 这项新技术将能够在制造和加工过程中定期监测蛋白质浓度。 一些抗体和其他蛋白质的市场每年在100亿美元的范围内，这些生物分子在治疗癌症、糖尿病和贫血等疾病方面至关重要。通过监测这些蛋白质的表达和加工来改善其生产是提高其可用性的重要步骤。 这项研究可能会产生一个简单的平台，用于大多数实验室使用的多孔板中的蛋白质分析。 考虑到治疗性蛋白质市场的规模（2500亿美元/年），用于分析治疗性蛋白质生产的工具应该具有每年数百万美元的市场。该项目将在96孔板中使用通过功能化微孔膜的流动，以在几分钟内捕获和检测蛋白质。 相比之下，典型的蛋白质分析平台依赖于被动扩散将蛋白质带到结合位点;分析时间为1小时。 这项研究包括一种用肽对多孔膜进行功能化的方法，这些肽可以特异性地与给定的蛋白质结合。 高密度的肽固定和膜中的高表面积与体积比允许在短时间内捕获大量蛋白质以实现快速、灵敏的定量。 该研究的目的是证明一个通用的方法，用于分析细胞培养液中的抗体，以及可比的分析时间，检测限，回收率和精度相比，商业免疫吸附测定。此外，这项工作可能建立基于膜的捕获和分析其他重要蛋白质，如胰岛素和促红细胞生成素。这些研究的成功可能证明功能化膜板用于定期监测重要商业蛋白质的生产和加工的前景。 该研究还将证明这种方法优于现有技术。该奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。

The broader impact/commercial potential of this Partnerships for Innovation - Technology Translation (PFI-TT) project includes development of a convenient device that can determine protein concentrations in 5 minutes. Current methods for protein analysis often employ expensive kits that require more than 1 hour to obtain results. The new technology will enable regular monitoring of protein concentrations during their manufacturing and processing. The markets for some individual antibodies as well as other proteins are in the range of $10 billion per year, and these biomolecules are vital in treating diseases such as cancer, diabetes, and anemia. Improving the production of these proteins by monitoring their expression and processing is an important step in enhancing their availability. The research may yield a simple platform for protein analysis in multi-well plates used in most laboratories. Given the size of the therapeutic protein market ($250 billion/year), tools for analysis of therapeutic protein production should have a market of many million dollars per year. This project will use flow through functionalized microporous membranes in 96-well plates to enable capture and detection of proteins in minutes. In contrast, typical protein analysis platforms rely on passive diffusion to bring proteins to binding sites; Analysis times are 1 h. The research includes a method for functionalizing the porous membranes with peptides that bind specifically to a given protein. The high density of peptide immobilization and the high surface area to volume ratio in the membranes allows capture of large amounts of protein in a short time to achieve rapid, sensitive quantitation. The research aims to demonstrate a general method for the analyses of antibodies in cell-culture broths as well as comparable analyses times, detection limits, percent recoveries, and precision when compared to commercial immunosorbent assays. Additionally, the work may establish membrane-based capture and analysis of other important proteins such as insulin and erythropoietin. Success in these studies may demonstrate the promise of functionalized membrane plates for regularly monitoring the production and processing of important commercial proteins. The research will also demonstrate the advantages of this method over current techniques.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.