Development of a droplet based microfluidic system for protein separation and fractionation

开发用于蛋白质分离和分级的基于液滴的微流体系统

• 批准号: 520804-2017
• 负责人: Ren, Carolyn
• 金额: $ 5.37万
• 依托单位: University of Waterloo
• 依托单位国家: 加拿大
• 项目类别: Collaborative Research and Development Grants
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=691700
• 关键词: Development; droplet; based; microfluidic; system

Capillary electrophoresis (CE) systems are important tools for the life sciences industry, particularly for protein analysis. Advanced Electrophoresis Solutions (AES) is an emerging leading developer of CE tools, and its flagship product, CEInfinite, provides reliable, high performance, and high throughput detection for protein separation and quantification using imaging detection capillary isoelectric focusing (iCIEF) technology. However, the company is facing critical challenges to meeting growing customer demand, and needs: i) low cartridge production rates due to slow, manual gluing processes; and ii) low purity and low volume of the recovered fraction of interest due to the use of single phase pressure driven flow (protein samples are mixed with buffer solution) for sample mobilization resulting in distorted sample bands and diluted protein fraction. Over the next three years, University of Waterloo researchers will work with AES to develop innovations that address the above issues as well as lay a foundation for AES to pursue future development in the field of single cell analysis (SCA), which holds tremendous potential for discovering biological mechanisms not seen in bulk solution settings. Specifically, we will: i) develop a two-phase droplet method, enabling sample injection with flexibly controlled volume; ii) develop a built-in membrane and a membrane-free strategy for the separation of electrolytes from protein sample mixtures required in iCIEF applications, and iii) develop a two-phase droplet method for continuous recovery of individual protein fractions with minimal dilution (high purity) and large volume. The proposed innovations will eliminate the need of gluing processes dramatically increasing cartridge production, and address the challenge of low volume and low purity of the recovered fraction of interest. The knowledge learned about active control of individual droplets will lay a foundation for AES to pursue future development in SCA as pico- to nano-liter droplets are ideal for isolation of single cells. The ultimate goal of this project is to help AES strongly penetrate into the pharmaceutical industry and also significantly expand its market into life science with emphasis on developing innovations and instruments for SCA.

毛细管电泳（CE）系统是生命科学行业的重要工具，尤其是用于蛋白质分析的重要工具。先进的电泳溶液（AES）是CE工具的新兴开发人员，其旗舰产品Ceinfinite使用成像检测毛细管等电向焦点（ICIEF）技术提供了可靠，高性能和高吞吐量检测，用于蛋白质分离和定量。但是，该公司面临满足不断增长的客户需求和需求的关键挑战：i）由于缓慢，手动胶合的过程而导致的墨盒生产率较低； ii）由于使用单相压力驱动流量（蛋白质样品与缓冲溶液混合）以进行样品动员，导致样品带和稀释的蛋白质分数导致样品动员，因此纯净和低体积的恢复率分数。在接下来的三年中，滑铁卢大学的研究人员将与AES合作开发解决上述问题的创新，并为AES在单细胞分析（SCA）领域追求未来发展的基础，该领域具有发现在散装解决方案环境中未见的生物学机制的巨大潜力。具体而言，我们将：i）开发一种两相液滴方法，使样品注入具有灵活控制的体积； ii）开发一种内置膜和一种无膜的策略，用于将电解质与ICIEF应用中所需的蛋白质样品混合物分离，iii）开发了一种两相液滴方法，用于连续恢复具有最小稀释度（高纯度）和较大体积的单个蛋白质分数。拟议的创新将消除胶合过程大大增加墨盒生产的需求，并应对恢复利益率的低量和低纯度的挑战。关于单个液滴积极控制的知识将为AES奠定基础，以追求SCA中的未来发展，因为PICO-纳米液滴是单个细胞隔离的理想选择。该项目的最终目的是帮助AES强烈渗透到制药行业，并将其市场大大扩展到生命科学，重点是开发SCA的创新和工具。