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

毛细管电泳（CE）系统是生命科学行业的重要工具，特别是用于蛋白质分析。Advanced Electrophoresis Solutions （AES）是一家新兴的领先的电泳工具开发商，其旗舰产品CEInfinite使用成像检测毛细管等电聚焦（iCIEF）技术，为蛋白质分离和定量提供可靠、高性能和高通量的检测。然而，该公司正面临着满足不断增长的客户需求的关键挑战，并且需要：i)由于缓慢的手动粘合过程导致墨盒生产率低；ii)由于使用单相压力驱动流（蛋白质样品与缓冲溶液混合）进行样品动员，导致样品带扭曲，蛋白质部分被稀释，因此回收率低，体积小。在接下来的三年里，滑铁卢大学的研究人员将与AES公司合作开发解决上述问题的创新技术，并为AES公司在单细胞分析（SCA）领域的未来发展奠定基础，该领域在发现大量溶液设置中未见的生物机制方面具有巨大的潜力。具体来说，我们将：i)开发一种两相液滴法，使进样能够灵活控制体积；ii)开发一种内置膜和无膜策略，用于从iCIEF应用所需的蛋白质样品混合物中分离电解质，iii)开发一种两相液滴法，用于以最小稀释（高纯度）和大体积连续回收单个蛋白质组分。提出的创新将消除粘接工艺的需要，大大增加了墨盒的产量，并解决了小体积和低纯度回收部分的挑战。所获得的关于单个液滴主动控制的知识将为AES在SCA领域的未来发展奠定基础，因为微米到纳升液滴是分离单细胞的理想选择。该项目的最终目标是帮助爱依斯公司大力渗透到制药行业，并将其市场扩展到生命科学领域，重点是为爱依斯公司开发创新和仪器。

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.