Institut de recherche en biotechnologie
6100, Royalmount, bureau L-334
T 514 496-6192
F 514 496-6785
- Expression and purification of recombinant proteins
- Large-scale transfection
- Cell culture in bioreactor
Dr Yves Durocher is a Project Leader in the Animal Cell Technology Group of the Biotechnology Research Institute of the National Research Council Canada. The Group consists of 30 researchers, engineers, technicians, students and post-doctoral fellows working on bioprocess developments using animal cells for the production of recombinant proteins (r-proteins) and viral vectors. The production of r-proteins is a critical step for elucidating their the role, function and structure. However, their production is often compromised by their post-translational complexity (glycosylation, disulfide bridge formation, proteolytic maturation, etc). Dr Durocher’s team has acquired an unique expertise in developing processes for the production of r-proteins using stable clones. In addition, his team has developed a large-scale transfection technology using HEK293 and CHO cells allowing the production of milligrams to grams quantities of r-proteins within 15 days following cloning of their cDNAs. This technology is used at up to the 45L-scale and has allow the production and purification of a vast array of secreted, membrane and cytosolic r-proteins. Dr Durocher’s work have shown that over 60% of the cells do express the gene of interest allowing the production of over 100 mg/L of r-proteins five days post-transfection. The research team currently works on further increasing the yields by improving the expression vectors, the cell lines, the culture medium composition and the transfection process. The team also investigate the mechanisms responsible for the capture and nuclear routing of the plasmids in order to develop new strategies to further improve the technology. The low costs, high yields and rapidity of protein expression provided by this technology offer significant advantages for both fundamental and industrial research.
- Boucher C., St-Laurent G., Loignon M., Jolicoeur M., De Crescenzo G., Durocher Y. (2008) The bioactivity and receptor affinity of recombinant tagged-EGF designed for tissue engineering applications is defined by the nature and position of the tags. Tissue Engineering, PMID: 18652537.
- S. Paris, A. Burlacu, and Y. Durocher (2008). Opposing roles of syndecan-1 and syndecan-2 in polyethylenimine-mediated gene delivery. Journal of Biological Chemistry, 283:7697-7704
- Tom, R., Bisson, L. and Durocher, Y. Transient expression in HEK293-EBNA1 cells. In M. Dyson and Y. Durocher, Editors. Methods Express Series – Expression Systems. Scion Publishing Ltd, Bloxham, Oxfordshire, UK, July 2007.
- Y. Durocher, P.L. Pham, G. St-Laurent, D. Jacob, B.Cass, P. Chahal, C. J. Lau, J. Nalbantoglu, A.Kamen. (2007) Serum-free production of recombinant adeno-associated virus type 2 by transfection of 293 suspension cells in bioreactors. Journal of Virological Methods, 144:32-40.
- Carpentier, E, Paris, S, Kamen A, Durocher Y. (2007) Limiting Factors Governing Protein Expression Following Polyethylenimine-Mediated Gene Transfer in HEK293-EBNA1 Cells. Journal of Biotechnology 128:268-80.
- Pham PL, Kamen A, Durocher Y. (2006) Large-Scale Transfection of Mammalian Cells for the Fast Production of Recombinant Protein. Molecular Biotechnology, 34:225-237.
- Shi C, Shin YO, Hanson J, Cass B, Loewen MC, Durocher Y. (2005) Purification and Characterization of a Recombinant G-Protein Coupled Receptor, Saccharomyces cerevisiae STE2p, Transiently Expressed in HEK293 EBNA1 Cells. Biochemistry, 44:15705-15714.
- Pham PL, Perret S, Cass B, Carpentier, E, St-Laurent G, Bisson L, Kamen A, Durocher Y. (2005) Transient Gene Expression in HEK293 Cells:Peptone Addition Posttransfection Improves Recombinant Protein Synthesis. Biotechnology and Bioengineering, 90:332-344.
- Cass B, Pham PL, Kamen A, Durocher Y. (2005) Purification of recombinant proteins from mammalian cell culture using a generic double affinity chromatography method. Protein Expression and Purification, 40:77-85.
- Durocher, Y., Perret, S. and Kamen, A. (2002) High Level and High-Throughput Recombinant Protein Production by Transient Transfection of the Suspension-Growing Human 293-EBNA1 Cell Line. Nucleic Acids Research, 30:e9.