Research Project

Website: http://www.oisb.ca/technology/technology_development.htm

Summary

Improvements in the protein analytical field are concerned primarily with increasing sensitivity. Where there have been vast performance improvements in recent years in techniques like mass spectrometry, these are not fully reflected in increased sensitivity because the samples are not efficiently processed prior to the analysis. This is one of the major challenges in the proteomics field. This project developed a novel technology, the proteomic reactor, based on the efficient handling of very small volumes of samples (microfluidics) with implications across the field of protein analysis, in particular studies of proteins in cells that appear in only trace levels in the body such as stem cells. This proteomic reactor simplifies handling of proteins, greatly reduces the volumes required for analysis and shortens processing times. It has provoked strong interest from both Canadian and international scientific equipment companies. 

 
The project’s objectives were to further improve and expand the proteomic reactor technology in two phases. In the first, the current format of the proteomic reactor was rapidly expanded, to four reactor column formats covering sample needs that range from large complex proteomes all the way down to a few cells. These systems were also made faster and more efficient with automated fluidic stations. By year two, automated proteomic reactor kits were developed and their novel applications in the field of rare cell analysis were demonstrated. In the second phase, the proteomic reactor was transposed and transformed using microfabrication technology available at the National Research Council. These microfluidic chips provided further enhancement in sample processing by reducing the reaction volumes, more efficient sample handling, and different surface biochemistry. Training sessions were done during year two of this proposal.

The proteomic reactor, in its different implementations, ranks as the most efficient proteomic processing device in terms of sensitivity, speed, and simplification of process. It was demonstrated that as few as 300 living cells can be processed and analyzed by mass spectrometry and that, even with higher cell numbers (such as 50,000 stem cells), the reactor provides better performance than conventional protein processing approaches. The tremendous potential of the multiplexed and automated proteomic reactor for the analysis of human embryonic stem cells and the analysis of very specific and minute regions of mouse and human brains was also demonstrated.

This project generated numerous positive outcomes, including different formats of the proteomic reactor that can handle various amounts of samples, more specialized reactors for the purification of specific subgroups of proteins, and an automated proteomic reactor that can automatically process and analyse complex proteomic samples. Novel surface chemistry and fabrication processes of the microfabricated proteomic reactor were also developed. As well, this project provided a unique cross-disciplinary training ground from human biology to engineering. At the international level, this project has facilitated the development of collaborations with groups in China, and facilitated the development of collaborative projects with industrial partners.

Notable Publications
Zhou, H., Hou, W., Denis, N.J., Zhou, H., Vasilescu, J., Zou, H., and Figeys, D.  2009.  Glycoproteomic Reactor for Human Plasma.  Journal of Proteome Research 8(2): 556-566