Production and Purification of Recombinant Proteins

  • Farida Kadir
  • Paul Ives
  • Alfred Luitjens
  • Emile van Corven


The growing therapeutic use of proteins has created an increasing need for practical and economical processing techniques. As a result, biotechnological production methods have advanced significantly over the last decade. Also, single-use production technology which has the potential to mitigate many of the economic and quality issues arising from manufacturing these products has evolved rapidly (Hodge 2004).


Recombinant Protein Affinity Chromatography Purification Process Downstream Processing Purification Scheme 
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  1. Afeyan N, Gordon N, Mazsaroff I, Varady L, Fulton S, Yang Y, Regnier F (1989) Flow-through particles of the high-performance liquid chromatographic separation of biomolecules, perfusion chromatography. J Chromatogr 519:1–29Google Scholar
  2. Benedek K, Swadesh JK (1991) HPLC of proteins and peptides in the pharmaceutical industry. In: Fong GW, Lam SK (eds) HPLC in the pharmaceutical industry. Dekker, New York, pp 241–302Google Scholar
  3. Berthold W, Walter J (1994) Protein purification: aspects of processes for pharmaceutical products. Biologicals 22:135–150PubMedCrossRefGoogle Scholar
  4. Borman S (2006) Glycosylation engineering. Chem Eng News 84:13–22Google Scholar
  5. Cartwright T (1987) Isolation and purification of products from animal cells. Trends Biotechnol 5:25–30CrossRefGoogle Scholar
  6. Celik E, Calik P (2011) Production of recombinant proteins by yeast cells. Biotechnol Adv. doi: 10.1016/j.biotechadv.2011.09.011 PubMedGoogle Scholar
  7. Chase HA (1994) Purification of proteins by adsorption chromatography in expanded beds. Trends Biotechnol 12:296–303PubMedCrossRefGoogle Scholar
  8. Chase H, Draeger N (1993) Affinity purification of proteins using expanded beds. J Chromatogr 597:129–145Google Scholar
  9. Compton B, Jensen J (2007) Use of perfusion technology on the Rise – New modes are beginning to gain ground on Fed-Batch strategy. Genetic Engineering & Biotechnology News 27(17):48Google Scholar
  10. Cumming DA (1991) Glycosylation of recombinant protein therapeutics: Control and functional implications. Glycobiology 1(2):115–130PubMedCrossRefGoogle Scholar
  11. Dell A, Galadari A, Sastre F, Hitchen P (2011) Similarities and differences in the glycosylation mechanisms in prokaryotes and eukaryotes. Int J Microbiol 2010(2010):148178Google Scholar
  12. EMA (2011) Note for guidance on minimising the risk of transmitting animal spongiform encephalopathy agents via human and veterinary medicinal products (EMA/410/01 rev.3)Google Scholar
  13. European Pharmacopoeia Seventh Edition. Strasbourg: Council of Europe (2011) 5.2.3. Cell substrates for the production of vaccines for human useGoogle Scholar
  14. FD3A, Center for Biologics Evaluation and Research (1990) Cytokine and growth factor pre-pivotal trial information package with special emphasis on products identified for consideration under 21 CFR 312 Subpart E. BethesdaGoogle Scholar
  15. FDA, Office of Biologicals Research and Review (1993) Points to consider in the characterization of cell lines used to produce biologicals. Rockville Pike/BethesdaGoogle Scholar
  16. Fulton SP (1994) Large scale processing of macromolecules. Curr Opin Biotechnol 5:201–205PubMedCrossRefGoogle Scholar
  17. Gottschalk U (2006) The renaissance of protein purification. BioPharm Int 19(6):S8–S9Google Scholar
  18. Heng M, Glatz C (1993) Charged fusions for selective recovery of ß-galactosidase from cell extract using hollow fiber ion-exchange membrane adsorption. Biotechnol Bioeng 42:333–338PubMedCrossRefGoogle Scholar
  19. Hjerten S, Mohammed J, Nakazato K (1993) Improvement in flow properties and pH stability of compressed, continuous polymer beds for high-performance liquid chromatography. J Chromatogr 646:121–128CrossRefGoogle Scholar
  20. Hodge G (2004) Disposable components enable a new approach to biopharmaceutical manufacturing. BioPharm Int 2004(15):38–49Google Scholar
  21. Homma T, Fuji M, Mori J, Kawakami T, Kuroda K, Taniguchi M (1993) Production of cellobiose by enzymatic hydrolysis: removal of ß-glucosidase from cellulase by affinity precipitation using chitosan. Biotechnol Bioeng 41:405–410PubMedCrossRefGoogle Scholar
  22. Horowitz MS, Bolmer SD, Horowitz B (1991) Elimination of disease-transmitting enveloped viruses from human blood plasma and mammalian cell culture products. Bioseparation 1:409–417Google Scholar
  23. ICH (International Conference on Harmonization) Topic Q6B (1999a) Specifications: test procedures and acceptance criteria for biotechnology/biological productsGoogle Scholar
  24. ICH (International Conference on Harmonization) Topic Q5A (1999b) Viral safety evaluation of biotechnology products derived from cell lines of human or animal originGoogle Scholar
  25. International Conference on Harmonization guideline M7 on assessment and control of DNA reactive (mutagenic) impurities in pharmaceuticals to limit potential carcinogenic riskGoogle Scholar
  26. James AM (1992) Introduction fundamental techniques. In: James AM (ed) Analysis of amino acids and nucleic acids. Butterworth-Heinemann, Oxford, pp 1–28Google Scholar
  27. Jones K (1990) Affinity chromatography, a technology up-date. Am Biotechnol Lab 8:26–30PubMedGoogle Scholar
  28. Kelley B (2009) Industrialization of mAb production technology. MAbs 1(5):443–452PubMedCrossRefGoogle Scholar
  29. Klegerman ME, Groves MJ (1992) Pharmaceutical biotechnology. Interpharm Press, Inc., Buffalo GroveGoogle Scholar
  30. Löwer J (1990) Risk of tumor induction in vivo by residual cellular DNA: quantitative considerations. J Med Virol 31:50–53PubMedCrossRefGoogle Scholar
  31. Maerz H, Hahn SO, Maassen A, Meisel H, Roggenbuck D, Sato T, Tanzmann H, Emmrich F, Marx U (1996) Improved removal of viruslike particles from purified monoclonal antibody IgM preparation via virus filtration. Nat Biotechnol 14:651–652PubMedCrossRefGoogle Scholar
  32. Marcus-Sekura CJ (1991) Validation and removal of human retroviruses. Center for Biologics Evaluation and Research, FDA, BethesdaGoogle Scholar
  33. Minor PD (1994) Ensuring safety and consistency in cell culture production processes: viral screening and inactivation. Trends Biotechnol 12:257–261PubMedCrossRefGoogle Scholar
  34. Monteclaro F (2010) Protein expression systems, ringing in the new. Innov Pharm Technol 12:45–49Google Scholar
  35. Note for Guidance (1991) Validation of virus removal and inactivation procedure, Ad Hoc Working Party on Biotechnology/Pharmacy, European Community, DG III/8115/89-ENGoogle Scholar
  36. Orzaez D, Granell A, Blazquez MA (2009) Manufacturing antibodies in the plant cell. Biotechnol J 4:1712–1724PubMedCrossRefGoogle Scholar
  37. PDA Journal of Pharmaceutical Science and Technology (2005) Technical report No. 41, Virus filtration, 59, No. S-2Google Scholar
  38. Peters J, Stoger E (2011) Transgenic crops for the production of recombinant vaccines and anti-microbial antibodies. Hum Vaccin 7(3):367–374PubMedCrossRefGoogle Scholar
  39. Sadana A (1989) Protein inactivation during downstream separation, part I: the processes. Biopharm 2:14–25Google Scholar
  40. Sharma SK (1990) Key issues in the purification and characterization of recombinant proteins for therapeutic use. Adv Drug Deliv Rev 4:87–111CrossRefGoogle Scholar
  41. Shukla AA, Thömmes J (2010) Recent advances in large-scale production of monoclonal antibodies and related proteins. Trends Biotechnol 28:253–261PubMedCrossRefGoogle Scholar
  42. Sinclair AM, Elliott S (2005) Glycoengineering: the effect of glycosylation on the properties of therapeutic proteins. J Pharm Sci 94:1626–1635PubMedCrossRefGoogle Scholar
  43. Tennikova T, Svec F (1993) High performance membrane chromatography: highly efficient separation method for proteins in ion-exchange, hydrophobic interaction and reversed phase modes. J Chromatogr 646:279–288CrossRefGoogle Scholar
  44. Terstappen G, Ramelmeier R, Kula M (1993) Protein partitioning in detergent-based aqueous two-phase systems. J Biotechnol 28:263–275PubMedCrossRefGoogle Scholar
  45. Van Wezel AL, van der Velden-de Groot CA, de Haan HH, van den Heuvel N, Schasfoort R (1985) Large scale animal cell cultivation for production of cellular biologicals. Dev Biol Stand 60:229–236PubMedGoogle Scholar
  46. Walsh C (2006) Posttranslational modification of proteins: expanding nature’s inventory, vol xxi. Roberts and Co. Publishers, Englewood, p 490Google Scholar
  47. Walter J, Werner RG (1993) Regulatory requirements and economic aspects in downstream processing of biotechnically engineered proteins for parenteral application as pharmaceuticals. In: Kroner KH, Papamichael N, Schütte H (eds) Downstream processing, recovery and purification of proteins, a handbook of principles and practice. Carl Hauser Verlag, MuenchenGoogle Scholar
  48. Walter J, Werz W, McGoff P, Werner RG, Berthold W (1991) Virus removal/inactivation in downstream processing. In: Spier RE, Griffiths JB, MacDonald C (eds) Animal cell technology: development, processes and products. Butterworth-Heinemann Ltd. Linacre House, Oxford, pp 624–634Google Scholar
  49. Walter K, Werz W, Berthold W (1992) Virus removal and inactivation, concept and data for process validation of downstream processing. Biotech Forum Europe 9:560–564Google Scholar
  50. Wheelwright SM (1993) Designing downstream processing for large scale protein purification. Biotechnology 5:789–793Google Scholar
  51. WHO (World Health Organization) (2010) Recommendations for the evaluation of animal cell cultures as substrates for the manufacture of biological medicinal products and for the characterization of cell banks. Technical report series, proposed replacement of 878, annex 1 (not yet published)Google Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Farida Kadir
    • 1
  • Paul Ives
    • 2
    • 3
  • Alfred Luitjens
    • 3
  • Emile van Corven
    • 3
  1. 1.Postacamedic Education Pharmacists (POA)BunnikThe Netherlands
  2. 2.Manufacturing DepartmentSynCo Bio Partners BVAmsterdamThe Netherlands
  3. 3.Department of Process DevelopmentCrucellLeidenThe Netherlands

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