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Process Economics of Animal Cell and Bacterial Fermentations: A Case Study Analysis of Tissue Plasminogen Activator

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Abstract

One link in the complex chain of medical economics is the cost of bringing new drugs and biologicals to the market. Advances in recombinant-DNA technology permit production of therapeutically active proteins in effectively unlimited quantities. Nevertheless, each expression system has a characteristic influence on the nature of the product produced and the process required to obtain it. In this case study we compare experiences with recombinant-tissue plasminogen activator (rtPA) produced in Chinese hamster ovary (CHO) cells and in Escherichia coli, with the aim of understanding the roles of some of the parameters that affect process economics. tPA belongs to the group of highly specific serine proteases that convert plasminogen to plasmin, which in turn degrades several protein substrates including fibrin, thus making it an effective thrombolytic agent. The treatment of acute myocardial infarction with such thrombolytic agents can result in early discharge of patients and decreased medical costs. However, there are major differences in the prices of the various available agents. The price of the FDA-licensed tPA product is $2,200 per dose or $22,000 per gram. It is believed that a significant portion of this price relates to manufacturing costs. We examine by way of case study illustration the cost breakdown for the two processes, and highlight important process, design and economic considerations that ultimately define a particular protein product.

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References

  1. 1

    Spalding, B.J. 1991. Downstream processing: Key to slashing production costs 100 fold. Bio/Technology 9: 229–233.

  2. 2

    Pennica, D., Holmes, W.E., Kohr, W.J., Harkins, R.N., Vehar, G.A., Ward, C.A., Bennet, W.F., Yelverton, E., Seeburg, P.H., Heyneker, H.L., Eodal, D.R. and Collen, D. 1983. Cloning and expression of tissue type plasminogen activator in E. coli. Nature 301: 214–221.

  3. 3

    Harris, T.J.R., Patel, T., Marston, F.A.O., Little, S., Emtage, S., Open-dakker, G., Volkaert, G., Rombauts, W., Billiau, A. and De Somer, P. 1986. Cloning of cDNA coding for human tPA and its expression in E. coli. Mol. Biol. Med. 3: 279–292.

  4. 4

    Thayer, A. 1991. Anti-clot drug study may hurt biotech sales. Chem. Eng. News. March 11: 6.

  5. 5

    Bluestone, M. 1992. tPA falls short once again. Bio/Technology 10: 480.

  6. 6

    Pohl, G., Kallstrom, M., Bergsdorf, N., Wallen, P. and Jornvall, H. 1984. Tissue plasminogen activator: peptide analyses confirm an indirectly derived amino acid sequence, identify the active site serine residue, establish glycosylation sites and localize variant differences. Biochemistry 23: 3701–3707.

  7. 7

    Collen, D., Stassen, J.M., Marafino, B.J. Jr., Builder, S., De Cock, F., Ogez, I., Tajiri, D., Pennica, D., Bennett, V.F., Salwa, J. and Hoyng, C.F. 1984. Biological properties of human tissue type plasminogen activator obtained by expression of recombinant DNA in animal cells. J. Pharmacol. Exp. Therap. 231: 146–152.

  8. 8

    Milstone, H. 1941. J. Immunol. 42: 109–116.

  9. 9

    Hansen, L., Blue, Y., Barone, K., Collen, D. and Larsen, G.R. 1988. Functional effects of asparagine-linked oligosaccharide on natural and variant human tissue-type plasminogen activator. J. Biol. Chem. 263: 15713–15719.

  10. 10

    Klausner, A. 1986. Researchers probe second-generation tPA. Bio/Technology 4: 706–711.

  11. 11

    Genentech, Inc. 1988. Third quarter report. September 30.

  12. 12

    Genentech, Inc. 1990. Activase® Info-Line. November.

  13. 13

    International reference standard tPA preparation (83/157). World Health Organization.

  14. 14

    Dodd, I., Jalapour, S., Southwick, W., Newsome, P., Browne, M.J. and Robinson, J.H. 1986. Large scale, rapid purification of recombinant tissue-type plasminogen activator. FEBS Lett. 209 13–17

  15. 15

    Schimke, R. 1978. Gene amplification in animal cells. Cell 37: 705–713.

  16. 16

    Stark, G. and Wahl, G. 1984. Gene amplification. Ann. Rev. Biochem. 53: 447–491.

  17. 17

    Wallen, P., Pohl, G., Bergsdorf, N., Ranby, M., Ny, T. and Jornall, H. 1983. Purification and characterization of a melanoma cell plasminogen activator. Bur. J. Biochem. 132: 681–686.

  18. 18

    Brouty-Boye, C.G., Maman, M., Marian, J. and Choay, P. 1984. Biosynthesis of human tissue-type plasminogen activator by normal cells. Bio/Technology 2: 1058–1062.

  19. 19

    Sarmientos, P., Duchesne, M., Denefte, P., Boiziau, J., Fromage, N., Delporte, N., Parker, F., Lelievre, Y., Mayaux, J.F. and Cartwright, T. 1989. Synthesis and purification of active human tissue plasminogen activator from E. coli. Bio/Technology 7: 495–501.

  20. 20

    Latta, M., Knapp, M., Sarmientos, P., Brefort, G., Becquart, J., Guerrier, L., Jung, G. and Mayaux, J.F. 1987. Synthesis and purification of native human serum albumin from E. coli. Bio/Technology 5: 1309–1314.

  21. 21

    Graff, R., Lang, K., Wrba, A. and Schmid, F.X. 1986. Folding mechanisms of porcine ribonuclease. J. Mol. Biol. 191: 281–293.

  22. 22

    Points to consider in the production and testing of new drugs and biologicals produced by recombinant DNA technology. 1985. US Office of Biologies Research and Review, FDA, Washington, DC.

  23. 23

    Datar, R. 1986. A comparative study of primary separation steps in fermentation. Ph.D. Thesis, Royal Institute of Technology, Stockholm, Sweden.

  24. 24

    Datar, R. and Rosen, C.-G. 1990. Downstream process economics, p. 741–793. In: Separation Processes in Biotechnology, Asenjo, J.A. (Ed.). Marcel Dekker,Inc., New York.

  25. 25

    Ulrich, G.D. 1984. A Guide to Chemical Engineering Process Design and Economics. John Wiley & Sons, Inc., New York.

  26. 26

    White, H.D., Rivers, J.T., Maslowski, A.H., Ormiston, J.A., Takayama, M., Hart, H.H., Sharpe, D.N., Whitelock, R.M.L. and Norris, R.M., 1989. Effect of intravenous streptokinase as compared with that of tissue plasminogen activator on left ventricular function after first myocardial infarction. N. Engl. J. Med. 320: 817–821.

  27. 27

    Grunfeld, H., Patel, A., Shatzman, A. and Nishikawa, A.H. 1992. Effector-assisted refolding of recombinant tissue-plasminogen activator produced in Escherickia coli. Appl. Biochem. Biotechnol. 33: 117–138.

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