Abstract
Therapeutic proteins have become a 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, composition, production, purification, contaminations, side effects, stability, formulation, and regulatory aspects.
This chapter describes basic elements of the selection, design, cloning, and production of biopharmaceuticals. The importance of the expression host in relation to specific posttranslational 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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Recommended reading and references
Agarwal KL, Büchi H, Caruthers MH, Gupta N, Korana HG, Kleppe K, Kumar A, Ohtsuka E, Rajbhandary UL, Van de Sande JH, Sgamarella V, Weber H, Yamada T (1970) Total synthesis of the gene for an alanine transfer ribonucleic acid from yeast. Nature 227(5253):27–34
Brekke OH, Sandlie I (2003) Therapeutic antibodies for human diseases at the dawn of the twenty-first century. Nat Rev Drug Discov 2(1):52–62
Carrara SC, Ulitzka M, Grzeschik J, Kornmann H, Hock B, Kolmar H (2021) From cell line development to the formulated drug product: the art of manufacturing therapeutic monoclonal antibodies. Int J Pharm 594:120164
Czar MJ, Anderson C, Bader JS, Peccoud J (2009) Gene synthesis demystified. Trends Biotechnol 27(2):63–72
EvaluatePharma® World Preview 2018(2018) Outlook to 2024 www.evaluate.com/PharmaWorldPreview2018 pdf (June 2018)
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. Microbiol Cell Fact 16:108
Gibson DG, Young L, Chuang RY, Venter JC, Hutchison CA, Smith HO (2009) Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods 6(5):343–345
Hartley JL (2003) Use of the gateway system for protein expression in multiple hosts. Curr Protoc Protein Sci:5.17.1–5.17.10. https://doi.org/10.1002/0471140864.ps0517s30
Hughes RA, Ellington AD (2017) Synthetic DNA synthesis and assembly: putting the synthetic in synthetic biology. Cold Spring Harb Perspect Biol 9(1):a023812
Kashmiri SV, De Pascalis R, Gonzales NR, Schlom J (2005) SDR grafting-a new approach to antibody humanization. Methods 36(1):2–34
Kim JY, Kim YG, Lee GM (2012) CHO cells in biotechnology for production of recombinant proteins: current state and further potential. Appl Microbiol Biotechnol 93:917–930
Kircher M, Kelso J (2010) High-throughput DNA sequencing–concepts and limitations. BioEssays 32(6):524–536
Kunert R, Reinhart D (2016) Advances in recombinant antibody manufacturing. Appl Microbiol Biotechnol 100:3451–3461
Lalonde ME, Durocher Y (2017) Therapeutic glycoprotein production in mammalian cells. J Biotechnol 251:128–140
Leader B, Baca QJ, Golan DE (2008) Protein therapeutics: a summary and pharmacological classification. Nat Rev Drug Discov 7(1):21–39
Lodish H, Berk A, Kaiser CA, Krieger M, Scott MP (2007) Molecular cell biology, 6th edn. WH. Freeman & CO, New York
Nothaft H, Szymanski CM (2010) Protein glycosylation in bacteria: sweeter than ever. Nat Rev Microbiol 8(11):765–778
Rosano GL, Morales ES, Ceccarelli EA (2019) New tools for recombinant protein production in Escherichia coli: a 5-year update. Protein Sci 28(8):1412–1422
SECRETERS-European Union’s Horizon 2020 Programme (2022) microbial protein cell factories fight back? Trends Biotechnol 40(5):576–590
Strohl WR, Knight DM (2009) Discovery and development of biopharmaceuticals: current issues. Curr Opin Biotechnol 20(6):668–672
Van Norman GA (2016) Drugs, devices, and the FDA: part 1: an overview of approval processes for drugs. JACC Basic Transl Sci 1(4):277–287
Wells EA, Robinson AS (2017) Cellular engineering for therapeutic protein production: product quality, host modification, and process improvement. Biotechnol J 12(16001015):1–12
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2024 The Author(s), under exclusive license to Springer Nature Switzerland AG
About this chapter
Cite this chapter
de Jong, O.G., Oosting, R.S. (2024). Molecular Biotechnology: From DNA Sequence to Therapeutic Protein. In: Crommelin, D.J.A., Sindelar, R.D., Meibohm, B. (eds) Pharmaceutical Biotechnology. Springer, Cham. https://doi.org/10.1007/978-3-031-30023-3_1
Download citation
DOI: https://doi.org/10.1007/978-3-031-30023-3_1
Published:
Publisher Name: Springer, Cham
Print ISBN: 978-3-031-30022-6
Online ISBN: 978-3-031-30023-3
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)