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Molecular Biotechnology: From DNA Sequence to Therapeutic Protein

  • Ronald S. OostingEmail author
Chapter

Abstract

Therapeutic proteins have become a very valuable addition to the repertoire of drugs to treat patients with severe diseases. Therapeutic proteins differ in many aspects from classical, small molecule drugs. They differ, for example, in molecular size, uniformity of the active, production, purification, contaminations, side effects, stability, formulation, regulatory aspects, and price.

This chapter describes basic elements of the selection, design, cloning and production of biopharmaceuticals. The importance of the expression host in relation to specific post-translational modifications, such as the formation of disulfide bridges and glycosylation, is discussed. And finally, we describe the molecular biological aspects of the design and production of (humanized) monoclonal antibodies in cell culture.

Keywords

Therapeutic protein Expression host Post-translational modification PCR Humanized monoclonal antibody 

Suggested Reading

  1. Brekke OH, Sandlie I (2003) Therapeutic antibodies for human diseases at the dawn of the twenty-first century. Nat Rev Drug Discov 2(1):52–62CrossRefGoogle Scholar
  2. EvaluatePharma® World Preview 2017 (2017) Outlook to 2022. http://info.evaluategroup.com/rs/607-YGS-364/images/WP17.pdf. Accessed 3 Apr 2018.
  3. GAciarz A, Khatri NK, Velez-Superbie ML, Saaranen MJ, Uchida Y, Keshavarz-Moore E, Ruddock LW (2017) Efficient soluble expression of disulfide bonded proteins in the cytoplasm of Escherichia coli in fed batch fermentations on chemically defined minimal media. Microbio Cell Fact 16:108CrossRefGoogle Scholar
  4. Kashmiri SV, De Pascalis R, Gonzales NR, Schlom J (2005) SDR grafting-a new approach to antibody humanization. Methods 36(1):2–34CrossRefGoogle Scholar
  5. Kim JY, kim Y-G, Lee GM (2012) CHO cells in biotechnology for production of recombinant proteins: current state and further potential. Appl Microbiol Biotechnol 93:917–930CrossRefGoogle Scholar
  6. Kircher M, Kelso J (2010) High-throughput DNA sequencing-concepts and limitations. Bioessays 32(6):524–536CrossRefGoogle Scholar
  7. Kunert R, Reinhart D (2016) Advances in recombinant antibody manufacturing. Appl Microbiol Biotechnol 100:3451–3461CrossRefGoogle Scholar
  8. Leader B, Baca QJ, Golan DE (2008) Protein therapeutics: a summary and pharmacological classification. Nat Rev Drug Discov 7(1):21–39CrossRefGoogle Scholar
  9. Lodish H, Berk A, Kaiser CA, Krieger M, Scott MP (2007) Molecular cell biology, 6th edn. WH. Freeman & CO, New YorkGoogle Scholar
  10. Nothaft H, Szymanski CM (2010) Protein glycosylation in bacteria: sweeter than ever. Nat Rev Microbiol 8(11):765–778CrossRefGoogle Scholar
  11. Paul SM, Mytelka DS, Dunwiddie CT, Persinger CC, Munos BH, Lindborg SR, Schacht AL (2010) How to improve R&D productivity: the pharmaceutical industry’s grand challenge. Nat Rev Drug Discov 9(3):203–214CrossRefGoogle Scholar
  12. Rosano GL, Ceccarelli EA (2014) Recombinant protein expression in Escherichia coli: advances and challenges. Front Microbiol 5(172):1–17Google Scholar
  13. Strohl WR, Knight DM (2009) Discovery and development of biopharmaceuticals: current issues. Curr Opin Biotechnol 20(6):668–672CrossRefGoogle Scholar
  14. Wells EA, Robinson AS (2017) Cellular engineering fro therapeutic protein production: product quality, host modification, and process improvement. Biotechnol J 12:16001015, 1–16001015,12CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Division of Pharmacology, Utrecht Institute for Pharmaceutical SciencesUtrecht UniversityUtrechtThe Netherlands

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