Abstract
The biotechnology industry today employs recombinant bacteria, mammalian cells, and transgenic animals for the production of high-value therapeutic proteins. This article reviews the techniques employed in this industry for the recovery of these products. The methods reviewed extend from the centrifugation and membrane filtration for the initial clarification of crude culture media to the final purification of the products by a variety of membrane-based and chromatographic methods. The subject of process validation including validation of the removal of bacterial and viral contaminants from the final products is also discussed with special reference to the latest regulatory guidelines.
Similar content being viewed by others
6. References
Lehrman, S.N. (2000) in Annual Report of Genzyme Transgenics, Framingham, MA., U.S.A.
Jack, K. (2001) Using transgenic cattle for the production of biologicals. Paper presented at the Conference on Production of Biopharmaceuticals (IBC Global Conferences), Hamburg, Germany.
Theisen, M. (2001) Production of therapeutic proteins in transgenic plants. Paper presented at the Conference on Production of Biopharmaceuticals (IBC Global Conferences), Hamburg, Germany.
Hamel, J-F.P. and Hunter, J.B. (1990). Modeling and applications of downstream processing — a survey of innovative strategies. in Downstream Processing and Bioseparation, ACS Symposium Series, (Hamel, J-F.P., Hunter, J.B. and Sikdar S.K., eds), ACS Chemical Society, Washington D.C., U.S.A.
Millipore Corporation (2000). Prefiltration and sterile filtration product selection guide: Optimised economics for biopharmaceutical applications. Lit.# SG 1 500EN00.
Valax, P, and Georgion, G. (1993). Molecular characterization of (-lactamase inclusion bodies produced in E coliM. 1. Composition., Biotechnol. Prog. 9, 539–547.
Middleberg, A.P.J. (2000). Microbial cell disruption by high pressure homogenization. in Methods in Biotechnology, Vol 9: Downstream Processing of Proteins: Methods and Protocols (M. A. Desai, Ed.) Pub. Humana Press Inc., Totowa, N.J.
Vogels G. and Kula, M.R. (1992). Combination of enzymatic and/or thermal pretreatment with mechanical cell disintegration. Chem. Eng. Sci. 47, 123–131.
Lutzer, R.G., Robinson, C.W., and Glick, B.R. (1994). Two stage process for increasing cell disruption of E. Coli for intracellular products recovery, in Proceedings of the 6th European Congress of Biotechnology, (Alberghina, A., Frontali, L and Sensi, P., eds). Elsevier Sciences B.V., Amsterdam, 111–121.
Baldwin, C. V. and Robinson, C.W. (1990). Disruption of Saccharomyces cerevisiae using enzymatic lysis combined with high pressure homogenization. Biotechnol. Tech., 4, 329–334.
Baldwin, C.V. and Robinson, C.W. (1994). Enhanced disruption of Candida utilis using enzymatic pretreatment and high pressure homogenization, Biotechnol. Bioeng., 43, 46–56.
Middleberg, A.P.J. (2000). Large scale recovery of protein inclusion bodies by continuous centrifugation in Methods in Biotechnology, Vol. 9: Downstream Processing of Proteins: Methods and Protocols. (Desai, M.A., ed.), Humana Press Inc., Totowa, NJ.
Strathman, H. (1985). Membranes and membrane processes in biotechnology, Trends in Biotechnol. 35, 112–118.
Belfort, G. (1987). in Advanced Biochemical Engineering (Bungay H.R. and Belfort, G., eds), Wiley and Sons, New York, NY, U.S.A.
Michaels, M., Michaels, A.S., Antoniou, C., et al. (1985). Tangential flow filtration. in Separations Technology: Pharmaceutical and Biotechnology Applications, (Olson, W.P., ed), Interpharm Press, Inc., Buffalo Grove, IL, U.S.A.
Vaheri, A., Gazzei, G. and Genna, G. (1981). Tangential flow filtration of Bordetella pertusis submerged cultures. Experentia, 35, 1535–1536.
Shibley, G.P., Manousas, M., Munch, K., Zelljadt, I., Fisher, L., and Mayyas, S. (1980). New method for large scale growth and concentration of Epstein Barr viruses. Appl. Environ. Microbiol. 40, 633–639.
Bellini, W.J., Trudgett, A., and McFarlin, D.E. (1979). Purification of measles virus with preservation of infectivity and antigenicity. J. Gen. Virol. 43, 633–639.
Zydney, A.L. and Kuriyel, R. (2000). Protein concentration and buffer exchange using ultrafiltration. in Methods in Biotechnology, Vol. 9: Downstream Processing of Proteins: Methods and Protocols, (M.A. Desai, ed.), Humana Press Inc., Totowa, NJ.
Wells, C.M., Lyddiatt, A. and Patel, K. (1987). Liquid fluidized bed adsorption in biochemical recovery from biological suspensions, in Separations for Biotechnol., (Verral, M.S. and Hudson, M.J., eds.) Ch. 16, 217–224, Ellis Horwood Ltd., Chichester, U.K.
Draeger, N.M. and Chase, H.A. (1991). Liquid fluidized bed adsorption of proteins in the presence of cells. Bioseparations, 2, 67–80.
McCormick, D.K. (1993). Expanded bed adsorption: The first new unit operation in decades. Bio/Technology, 11, 1059.
Chase, H.A. (1994). Purification of proteins by adsorption chromatography in expanded beds. TIBTECH, 12, 296–303.
Lihme, A., Hansen, M., Olander, M. and Zafirakos, E. (2000). Expanded bed adsorption in the purification of biomolecules, in Methods in Biotechnology, Vol. 9: Downstream Processing of Proteins: Methods and Protocols, (Desai, M.A., ed.), Humana Press Inc., Totowa, NJ.
Desai, M.A., Rayner, M., Burns, M. and Bermingham, D. (2000). Applications of chromatography in down-stream processing of biomolecules. in Methods in Biotechnology, Vol. 9: Downstream Processing of Proteins: Methods and Protocols, (Desai, M.A., ed.), Humana Press Inc., Totowa, NJ.
Mazek, J. and Kypr, J. (1992). Nucleotide composition of genes and hydrophobicity of the encoded proteins. FEBS Lett. 305, 163–165.
Eriksson, K. (1989). in Protein purification (Rydén, J., ed.), VCH, New York, NY.
Hearn, M.T.W. (1989). in Protein purification (Rydén, J., ed), VCH, New York, NY.
Manzke, O., Tesch, H., Diehl, V. and Bohlen, H. (1997). Single step purification of biospecific monoclonal antibodies for immunotherapeutic use by hydrophobic interaction chromatography. J. Immunol. Methods, 208, 65–73.
Gel Filtration: Principles and Methods, 6th edition, Pharmacia Biotech., Uppsala, Sweden.
Desai, M.A. (2000). Immunoaffinity adsorption in the large scale isolation of biomolecules, in Methods in Biotechnology, Vol. 9: Downstream Processing of Proteins: Methods and Protocols, (Desai, M.A., ed.) Humana Press, Inc., Totowa, NJ.
Walters, R. (1985). Affinity Chromatography. Anal. Chem. 57, 1099A-1114A.
Jervis, L. (1987). Polymers in affinity chromatography in Developments in Polymer Reagents, Catalysts and Adsorbents (Hodge, P. and Sherringtoon, D.C., eds.), Wiley, London, U.K., pp.65–96.
Sada, E., Katoh, S., Sukai, K., Tohma, M. and Kondo, A. (1986). Adsorption equilibrium in immunoaffinity chromatography with polyclonal and monoclonal antibodies. Biotechnol. Bioeng. 28, 1497–1502.
Desai, M.A. (1990) Immunoaffinity adsorption: Process scale isolation of therapeutic grade biochemicals. J. Technol. Biotechnol. 48, 105–106.
Nelsen, L.N. (1978) Removal of pyrogens from parenteral solutions by ultrafiltration. Pharma. Technol. 2(5), 46–49, 80.
Gabler, F.R. (1987). Pyrogens and the depyrogenation of solutions with ultrafiltration membranes. in Filtration in the Pharmaceutical Industry, (Meltzer, T., Ed.), Marcel Dekker, New York, NY.
Abramson, D., Butler, D. and Chrai, S. (1981). Depyrogenation of a parenteral solution by ultrafiltration. J. Parent. Sci. Technol. 35 (1), 3–7.
Kalyanpur, M., (2000). Validation of sterilising filters in the biotechnology industry, in Methods in Biotechnology, Vol. 9: Downstream Processing of Proteins: Methods and Protocols, (Desai, M.A., ed.), Humana Press, Inc., Totowa, NJ.
U.S. Food and Drug Administration (1987). Guideline on Sterile Drug Products Produced by Aseptic Processing, Center for Drugs and Biologics, Rockville, MD, U.S.A.
ICH Topic Q 5 A: Quality of Biotechnological Products — Viral Safety Evaluation of Biotechnology Products Derived from Cell Lines of Human or Animal Origin, European Agency of the Evaluation of Medicinal Products, Human Medicines Evaluation Unit, (CPMP/ICH/295/95), March 1997.
Burnouf-Radosevich, M., Appourchaux, P., Huart, J.J. and Burnhouf, T. (1994). Nanofiltration, a new specific virus elimination method applied to high purity Factor IX and Factor IX concentrates. Vox Sang 67, 132–138.
Hoffer, L., Schwinn, H., Biesert, H., and Josic, D. (1995). Improved virus safety and purity of a chromatographically produced Factor IX concentrate by nanofiltration. J. Chromatogr. 669, 187–196.
DiLeo, A.J., Vacante, D.A., and Dean, E.F. (1993a). Size exclusion removal of model mammalian viruses using a unique membrane system, Part I: Membrane qualification. Biologicals, 21, 275–286.
DiLeo, A.J., Vacante D.A., and Dean, E.F. (1993b). Size exclusion removal of model mammalian viruses using a unique membrane system, Part II: Module qualification and process simulation. Biologicals, 21, 287–296.
Levy, R.V., Phillips, M., and Lutz, H. (1998) Filtration and removal of viruses from biopharmaceuticals, in Filtration in the Biopharmaceutical Industry, (Meltzer, T.H. and Jorintz, M.W., eds.), Marcel Dekker, New York, NY.
Darling, A.J. (2000). Design and interpretation of Viral Clearance Studies for Biopharmaceutical Products, in Methods in Biotechnology, Vol. 9: Downstream Processing of Proteins: Methods and Protocols (Desai, M.A., ed) Humana Press Inc., Totowa, NJ.
U. S. Food and Drug Administration, Center for Drugs, Biologics, Devices and Radiologic Health (1987). Guidelines on general principles of process validations, Rockville, MD.
Commission of the European Communities (1989). Guide to good manufacturing of medicinal products, Vol. IV. The rules governing medicinal products in the European community, Luxembourg, Office for Official Publications of the European Communities.
Michaels, S.L. (1991). Validation of tangential flow filtration systems. J. Parent. Sci. and Technol. 45 (5), 218–223.
Millipore Corporation, Bedford, MA, U.S.A. (1990). Validation of tangential flow filtration (TFF) systems, lit. # TB 052.
Parenteral Drug Association (1992). Industry perspective on the validation of column-based separation processes for the purification of proteins. J. Parent. Sci. and Technol. 46, 87–97.
Kuwahara, S.S. and Chuan, J.H. (1995). Process validation of separation systems in Separations Technology: Pharmaceutical and Biotechnology Applications, (Olson, W.P., ed.), Interpharm Press, Inc., Buffalo Grove, IL.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kalyanpur, M. Downstream processing in the biotechnology industry. Mol Biotechnol 22, 87–98 (2002). https://doi.org/10.1385/MB:22:1:087
Issue Date:
DOI: https://doi.org/10.1385/MB:22:1:087