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Protein Downstream Processing pp 3–10Cite as

Protein Purification: An Overview

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1129))

Abstract

Biological macromolecules such as proteins constitute an important class of products in the food, biotechnology, pharmaceutical, and cosmetics industries. The growing need to develop efficient and rapid protein purification methods is driving research and growth in this area. Advances and progress in the methods and techniques of protein purification have been such that one can reasonably expect that any protein of a given order of stability may be purified to currently acceptable standards of homogeneity. However, protein production cost remains extremely high, with downstream processing constituting a substantial proportion of the overall cost. Understanding of the methods and optimization of experimental conditions have become critical to the manufacturing industry in order to minimize production costs while satisfying all regulatory requirements. New purification protocols exploiting specific, effective, and robust methods and materials are expected to guide the future of the protein purification area.

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References

  1. Shendure J, Lieberman AE (2012) The expanding scope of DNA sequencing. Nat Biotechnol 30:1084–1094

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  2. Henry RJ, Edwards M, Waters DL et al (2012) Application of large-scale sequencing to marker discovery in plants. J Biosci 37:829–841

    Article  CAS  PubMed  Google Scholar 

  3. Vaudel M, Sickmann A, Martens L (2012) Current methods for global proteome identification. Expert Rev Proteomics 9:519–532

    Article  CAS  PubMed  Google Scholar 

  4. Marichal-Gallardo PA, Alvarez MM (2012) State-of-the-art in downstream processing of monoclonal antibodies: process trends in design and validation. Biotechnol Prog 28:899–916

    Article  CAS  PubMed  Google Scholar 

  5. Kalyanpur M (2002) Downstream processing in the biotechnology industry. Mol Biotechnol 22:87–98

    Article  CAS  PubMed  Google Scholar 

  6. Kallberg K, Johansson HO, Bulow L (2012) Multimodal chromatography: an efficient tool in downstream processing of proteins. Biotechnol J 7:1485–1495

    Article  CAS  PubMed  Google Scholar 

  7. Cohn EJ, Edsall JT (1943) Proteins, amino acids and peptides as ions and dipolar ions. Reinhold Publishing, New York, NY

    Google Scholar 

  8. Lucy CA (2003) Evolution of ion-exchange: from Moses to the Manhattan Project to modern times. J Chromatogr A 1000:711–724

    Article  CAS  PubMed  Google Scholar 

  9. Starkenstein E (1910) Ferment action and the influence upon it of neutral salts. Biochem Z 24:210–218

    CAS  Google Scholar 

  10. Cuatrecasas P, Wilcheck M, Anfinsen CB (1968) Selective enzyme purification by affinity chromatography. Proc Natl Acad Sci U S A 61:636–643

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  11. Butler M, Meneses-Acosta A (2012) Recent advances in technology supporting biopharmaceutical production from mammalian cells. Appl Microbiol Biotechnol 96:885–894

    Article  CAS  PubMed  Google Scholar 

  12. Wilken LR, Nikolov ZL (2012) Recovery and purification of plant-made recombinant proteins. Biotechnol Adv 30:419–433

    Article  CAS  PubMed  Google Scholar 

  13. Rosa PA, Azevedo AM, Sommerfeld S et al (2011) Aqueous two-phase extraction as a platform in the biomanufacturing industry: economical and environmental sustainability. Biotechnol Adv 29:559–567

    Article  CAS  PubMed  Google Scholar 

  14. Chon JH, Zarbis-Papastoitsis G (2011) Advances in the production and downstream processing of antibodies. Nat Biotechnol 28:458–463

    CAS  Google Scholar 

  15. Freitag R, Horváth C (1996) Chromatography in the downstream processing of biotechnological products. Adv Biochem Eng Biotechnol 53:17–59

    CAS  PubMed  Google Scholar 

  16. Roque AC, Silva CS, Taipa MA (2007) Affinity-based methodologies and ligands for antibody purification: advances and perspectives. J Chromatogr A 1160:44–55

    Article  CAS  PubMed  Google Scholar 

  17. Labrou NE (2003) Design and selection of ligands for affinity chromatography. J Chromatogr B Analyt Technol Biomed Life Sci 790:67–78

    Article  CAS  PubMed  Google Scholar 

  18. Clonis YD (2006) Affinity chromatography matures as bioinformatic and combinatorial tools develop. J Chromatogr A 1101:1–24

    Article  CAS  PubMed  Google Scholar 

  19. Gottschalk U (2008) Bioseparation in antibody manufacturing: the good, the bad and the ugly. Biotechnol Prog 24:496–503

    Article  CAS  PubMed  Google Scholar 

  20. ISO 14644-1 (1999) Cleanrooms and associated controlled environments-part 1: classification of air cleanliness cleanroom standards

    Google Scholar 

  21. ISPE (2009) Baseline pharmaceutical engineering guides for new and renovated facilities. In: Biopharmaceuticals, vol 6. ISPE, Tampa, Fl, Glossary updated 06-23-2010

    Google Scholar 

  22. Rasmussen SK, Næsted H, Müller C et al (2012) Recombinant antibody mixtures: production strategies and cost considerations. Arch Biochem Biophys 526:139–145

    Article  CAS  PubMed  Google Scholar 

  23. Kelley B (2009) Industrialization of mAb production technology: the bioprocessing industry at a crossroads. MAbs 5:443–452

    Article  Google Scholar 

  24. Dranitsaris G, Amir E, Dorward K (2011) Biosimilars of biological drug therapies: regulatory, clinical and commercial considerations. Drugs 71:1527–1536

    Article  CAS  PubMed  Google Scholar 

  25. Davies HM (2010) Commercialization of whole-plant systems for biomanufacturing of protein products: evolution and prospects. Plant Biotechnol J 8:845–861

    Article  PubMed  Google Scholar 

  26. Huang CJ, Lin H, Yang X (2012) Industrial production of recombinant therapeutics in Escherichia coli and its recent advancements. J Ind Microbiol Biotechnol 39:383–399

    Article  CAS  PubMed  Google Scholar 

  27. Chen R (2012) Bacterial expression systems for recombinant protein production: E. coli and beyond. Biotechnol Adv 30:1102–1107

    Article  CAS  PubMed  Google Scholar 

  28. Young CL, Britton ZT, Robinson AS (2012) Recombinant protein expression and purification: a comprehensive review of affinity tags and microbial applications. Biotechnol J 7:620–634

    Article  CAS  PubMed  Google Scholar 

  29. Protein Engineering Market [Products (monoclonal antibody, insulin analog, modified EPO), technology (sequential modification, glycosylation, pegylation), and applications (therapeutics, diagnostics, research)] – Global Forecast to 2017

    Google Scholar 

  30. Walsh G (2010) Biopharmaceutical benchmarks 2010. Nat Biotechnol 28:917–924

    Article  CAS  PubMed  Google Scholar 

  31. Global Biosimilars Market Forecast to 2015. http://www.reportlinker.com/p0795429-summary/Global-Biosimilars-Market-Forecast-to.html

  32. Bird C (2011) Protein purification reagent market expands, role in biopharmaceutical, diagnostic, and basic biomedical research increases usage. Genetic engineering and biotechnology news, 31. http://www.genengnews.com/gen-articles/protein-purification-reagent-market-expands/3930/

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Acknowledgements

This work was supported by the grants: AlgaeCom and AquaPhage (funded by the European Union, FP7), BioExplore (09ΣΥΝ-23-94, co-funded by the European Union—European Social Fund & National Resources), and THALES (MIS380236, co-funded by the European Union—European Social Fund & National Resources).

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Authors and Affiliations

  1. Enzyme Technology Laboratory, Department of Biotechnology, Agricultural University of Athens, Iera Odos 75, 11855, Athens, Greece

    Nikolaos E. Labrou

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  1. Nikolaos E. Labrou
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Correspondence to Nikolaos E. Labrou .

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Editors and Affiliations

  1. Laboratory of Enzyme Technology, Agricultural University of Athens Department of Agr. Biotechnology, Athens, Greece

    Nikolaos E. Labrou

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Labrou, N.E. (2014). Protein Purification: An Overview. In: Labrou, N. (eds) Protein Downstream Processing. Methods in Molecular Biology, vol 1129. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-977-2_1

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  • DOI: https://doi.org/10.1007/978-1-62703-977-2_1

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-976-5

  • Online ISBN: 978-1-62703-977-2

  • eBook Packages: Springer Protocols

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