Simulation of large-scale production of a soluble recombinant protein expressed in Escherichia coli using an intein-mediated purification system
Inteins are self-cleavalbe proteins that under reducing conditions can be cleaved from a recombinant target protein. Industrially, an intein-based system could potentially reduce production costs of recombinant proteins by facilitating a highly selective affinity purification using an inexpensive substrate such as chitin. In this study, SuperPro® Designer was used to simulate the large-scale recovery of a soluble recombinant protein expressed in Escherichia coli using an intein-mediated purification process based on the commercially available IMPACT® system. The intein process was also compared with a conventional process simulated by SuperPro. The intein purification process initially simulated was significantly more expensive than the conventional process, primarily owing to the properties of the chitin resin and high reducing-agent (dithiothreitol [DTT]) raw material cost. The intein process was sensitive to the chitin resin binding capacity, cleavage efficiency of the intein fusion protein, the size of the target protein relative to the intein tag, and DTT costs. An optimized intein purification process considerably reduced costs by simulating an improved chitin resin and alternative reduced agents. Thus, to realize the full potential of intein purification processes, research is needed to improve the properties of chitin resin and to find alternative, inexpensive raw materials.
Index EntriesChitin process simulation recombinant protein production intein economic analysis
Unable to display preview. Download preview PDF.
- 1.Petrides, D. P., Koulouris, A., and Lagonikos, P. T. (2002), Pharm. Eng. 22, 56–65.Google Scholar
- 19.Ladisch, M. (2001), Bioseparations Engineering: Principles, Practice, and Economics, 1st ed., John Wiley & Sons, Indianapolis.Google Scholar
- 20.Protein Purification—Handbook. (2001), Amersham Biosciences AB, Uppsala, Sweden.Google Scholar
- 25.Brierley, R. A., Abrams, J. N., Hanson, J. M., and Maslanka, F. C. (2001), US patent 6,207,806 B1.Google Scholar
- 26.Koths, K. Thomson, J., Kunitani, M., Wilson, K., and Hanisch, W. (1986), US patent 4,569,790.Google Scholar
- 27.Stern, A. S. (1998), US patent 5,831,022.Google Scholar
- 29.Harrison, R. G., Todd, P., Rudge, S. R., and Petrides, D. P. (2003), Bioseparations Science and Engieering, Oxford University Press, New York.Google Scholar
- 30.Peters, M. S. and Timmerhaus, K. D. (1991), Plant Design and Economics for Chemical Engineers, 4th ed., McGraw-Hill, New York.Google Scholar
- 31.Garnett, D. I. and Patience, G. S. (1993), Chem. Eng. Prog. 89, 76–78.Google Scholar
- 32.Roberts, G. A. F. and Taylor, K. E. (1988), in Chitin and Chitosan—Sources, Chemistry, Biochemistry, Physical Properties and Applications, Skjak-Braek, G., Anthonsen, T., and Sandford, P., eds., Elsevier Applied Science, London, pp. 577–583.Google Scholar
- 33.Seo, H. and Kinemura, Y. (1988), in Chitin and Chitosan—Sources, Chemistry, Biochemistry, Physical Properties and Applications, Skjak-Braek, G., Anthonsen, T., and Sandford, P., eds., Elsevier Applied Science, London, pp. 585–588.Google Scholar