The Gene and Linda Voiland School of

Chemical Engineering and Bioengineering

Faculty

James M. Lee, Ph.D.

Professor

The Gene and Linda Voiland
School of Chemical Engineering and Bioengineering
Washington State University
118 Dana Hall Spokane St.
P.O. Box 642710
Pullman, WA 99164-2710



Office: EME B55 (Tel: 509 335-5252)
jmlee@wsu.edu
Dr. Lee's Website

Graduate Students

Rigoberto Rios-Estepa

Research Interests

Transgenic plant cell suspension cultures can be utilized for the production and secretion of biologically active mammalian proteins. Plant cell cultures may offer an inexpensive alternative to the mammalian system for the production of valuable foreign proteins. However, several difficulties remain before this promising new protein production technology can be commercially viable. Primary among these is the fact that the level of foreign protein expression within transgenic plant cells has to be significantly increased. Furthermore, more efficient fermenter systems must be developed to produce and recover the secreted mammalian proteins. The objective of our research is to enhance the expression, secretion, and recovery of mammalian proteins from transgenic plant suspension cultures. Various human lymphokines including monoclonal antibodies, GM-CSF, and interleukins (IL-2, IL-4, and IL-10) have been employed as model transgenic protein products. The expression of mammalian proteins in plant cells could be increased by using strong, constitutive and inducible promoters, or by adding the non-translated leader sequence of the tobacco etch virus (TEV). The targeting and secretion of transgenic proteins was examined by adding various plant signal peptide sequences. Protein recovery was maximized through the testing of a variety of additives for their effectiveness in stabilizing secreted transgenic proteins. A novel affinity-chromatography bioreactor was developed that uses the nickel-chelate nitrilotriacetic column to increase in situ production and separation of the secreted mammalian protein from plant suspension cultures.

This is an interdisciplinary research project (funded by National Science Foundation) with Drs. Nancy Magnuson and Raymond Reeves in Molecular Biosciences.

Selected Publications

  1. James, E. A., Lee, J. M. "Loss and Recovery of Protein Productivity in Genetically Modified Plant Cell Line," Plant Cell Reports, In Press (2006)
  2. Soderquist, R. G., Lee, J. M. "Enhanced Production of Recombinant Proteins from Plant Cells by the Application of Osmotic Stress," Plant Cell Reports, 24, 127-132 (2005)
  3. James, E. A., Mills, D. R., Lee, J. M. "Increased Production and Recovery of Foreign Proteins from Plant Cell Cultures Using an Affinity Chromatography Bioreactor," Biochem. Eng. J., Biochem. Eng. J., 12, 205-213 (2002)
  4. Magnuson, N. S., Wang, Z., James, E. A., Reeves, R., Lee, J. M. "Gene Cloning and Expression Technologies Plant cell culture expression systems," In M. Weiner and Q. Lu (editors), Gene Cloning and Expression Technologies, In Press, (2002)
  5. Xu, H., Montoya, F. U., Wang, Z., Lee, J. M., Reeves, R., Linthicum, D. S., Magnuson, N. S. "Combined use of regulatory elements within the cDNA to increase the production of a soluble mouse single chain antibody, scFv, from tobacco cell suspension cultures," Protein Exp. Pur., 24, 384-394, (2002)
  6. James, E. A., Mills, D. R., Lee, J. M. "Increased Production and Recovery of Foreign Proteins from Plant Cell Cultures Using an Affinity Chromatography Bioreactor," Biochem. Eng. J., In Press (2002)
  7. Magnuson, N. S., Wang, Z., James, E. A., Reeves, R., Lee, J. M. "Gene Cloning and Expression Technologies Plant cell culture expression systems," In M. Weiner and Q. Lu (editors), Gene Cloning and Expression Technologies, In Press, (2002)
  8. Xu, H., Montoya, F. U., Wang, Z., Lee, J. M., Reeves, R., Linthicum, D. S., Magnuson, N. S. "Combined use of regulatory elements within the cDNA to increase the production of a soluble mouse single chain antibody, scFv, from tobacco cell suspension cultures," Protein Exp. Pur., 24, 384-394, (2002)
  9. Bodeutsch, T., James, E. A., Lee, J. M. "The Effect of Immobilization on Recombinant Protein Production in Plant Cell Culture" Plant Cell Report, 20, 562-566, (2001)
  10. James, E. A., Lee, J. M. "The Production of Foreign Protein from Genetically Modified Plant Cells" In: J. J. Zhong (editor) Advances in Biochemical Engineering/Biotechnology - Plant Cells. Vol 72, pp 127-156, Springer, Berlin, Germany, (2001)
  11. Bodeutsch, T., James, E. A., Lee, J. M. "The Effect of Immobilization on Recombinant Protein Production in Plant Cell Culture" Plant Cell Report, 20, 562-566, (2001)
  12. Ryland, J. R., Linzmaier, P. M., Lee, J. M. "Effect of Gelatin on the Stability of Heavy Chain Monoclonal Antibody Production from Plant Suspension Cultures," J. Micro. Biotech., 10, 449-454, (2000)
  13. Ryland, J. R., Linzmaier, P. M., Lee, J. M. "Effect of Gelatin on the Stability of Heavy Chain Monoclonal Antibody Production from Plant Suspension Cultures," J. Micro. Biotech., 10, 449-454, (2000)

The Gene & Linda Voiland School of Chemical Engineering and Bioengineering, PO Box 642710, Washington State University, Pullman WA 99164-2710, 509-335-4332, Email ChEBE: chebe AT wsu DOT edu