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

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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.