Protein Production for Biotechnology

Peter J Korajczyk, Argonne National Laboratory, Lemont, Illinois, USA
Sarah Zerbs, Argonne National Laboratory, Lemont, Illinois, USA
Frank R Collart, Argonne National Laboratory, Lemont, Illinois, USA

Published online: October 2017

DOI: 10.1002/9780470015902.a0000983.pub3

Abstract

Purified proteins are useful as experimental tools, commercial products and therapeutics for the treatment of disease. The benefits of these products are dependent on specific qualities such as protein functionality, yield and purity. To meet these protein production criteria, a vast repertoire of constructs, genetic tools, methods and host organisms has been developed to enable multiple approaches for the production of diverse protein products. Consideration of the advantages and limitations of various heterologous expression systems is necessary to ensure that the resulting proteins will be useful for the specific needs of the intended project. Multiple approaches are often evaluated to accommodate the wide diversity in protein characteristics and functions. A systematic approach is outlined for evaluation of protein properties incorporating the requirements for the intended use of protein production to provide a reliable strategy for the selection of appropriate system(s) for large‐scale protein production.

Key Concepts

  • Controlled production of purified proteins is necessary for scientific research and has many therapeutic and commercial applications.
  • The intended use of a protein product strongly influences the methods selected for protein production.
  • Each biological expression system has unique advantages, disadvantages and requirements for protein production.
  • Multiple technological approaches and hosts may be screened to meet project requirements.
  • In vitro expression methods allow for the systematic development of proteins with unique modifications or properties.
  • A large resource of technological materials in support of protein production enables the generation of products with substantial economic and scientific interest.

Keywords: recombinant protein expression; fusion tag; cell‐free; biologics; genetic engineering; protein characterisation; chemical protein synthesis

Figure 1. Overview of heterologous protein production.
Figure 2. Automation platform for high‐throughput protein production.
close

References

Alberti F, Foster GD and Bailey AM (2017) Natural products from filamentous fungi and production by heterologous expression. Applied Microbiology and Biotechnology 101: 493–500.

Angius F, Ilioaia O, Uzan M and Miroux B (2016) Membrane protein production in Escherichia coli: protocols and rules. Methods in Molecular Biology 1432: 37–52.

Behrendt R, White P and Offer J (2016) Advances in Fmoc solid‐phase peptide synthesis. Journal of Peptide Science 22: 4–27.

Biedendieck R (2016) A Bacillus megaterium system for the production of recombinant proteins and protein complexes. Advances in Experimental Medicine and Biology 896: 97–113.

Cappuccio JA, Blanchette CD, Sulchek TA, et al. (2008) Cell‐free co‐expression of functional membrane proteins and apolipoprotein, forming soluble nanolipoprotein particles. Molecular and Cellular Proteomics 7: 2246–2253.

Dawson PE and Kent SB (2000) Synthesis of native proteins by chemical ligation. Annual Review of Biochemistry 69: 923–960.

Druzinec D, Salzig D, Brix A, et al. (2013) Optimization of insect cell based protein production processes ‐ online monitoring, expression systems, scale up. Advances in Biochemical Engineering and Biotechnology 136: 65–100.

Dumont J, Euwart D, Mei B, Estes S and Kshirsagar R (2016) Human cell lines for biopharmaceutical manufacturing: history, status, and future perspectives. Critical Reviews in Biotechnology 36: 1110–1122.

Gustafsson C, Minshull J, Govindarajan S, et al. (2012) Engineering genes for predictable protein expression. Protein Expression and Purification 83: 37–46.

Houdebine LM (2000) Transgenic animal bioreactors. Transgenic Research 9: 305–320.

Houdebine LM (2007) Transgenic animal models in biomedical research. Methods in Molecular Biology 360: 163–202.

Jia B and Jeon CO (2016) High‐throughput recombinant protein expression in Escherichia coli: current status and future perspectives. Open Biology 6.

Katzen F, Chang G and Kudlicki W (2005) The past, present and future of cell‐free protein synthesis. Trends in Biotechnology 23: 150–156.

Katzen F, Fletcher JE, Yang JP, et al. (2008) Cell‐free protein expression of membrane proteins using nanolipoprotein particles. Biotechniques 45: 469.

Kimple ME, Brill AL and Pasker RL (2013) Overview of affinity tags for protein purification. Current Protocols in Protein Science 73: Unit 9.9.

Kost TA, Condreay JP and Jarvis DL (2005) Baculovirus as versatile vectors for protein expression in insect and mammalian cells. Nature Biotechnology 23: 567–575.

Kost TA and Kemp CW (2016) Fundamentals of baculovirus expression and applications. Advances in Experimental Medicine and Biology 896: 187–197.

Laible PD, Scott HN, Henry L and Hanson DK (2004) Towards higher‐throughput membrane protein production for structural genomics initiatives. Journal of Structural and Functional Genomics 5: 167–172.

Lakowitz A, Godard T, Biedendieck R and Krull R (2017) Mini review: recombinant production of tailored bio‐pharmaceuticals in different Bacillus strains and future perspectives. European Journal of Pharmaceutics and Biopharmaceutics. pii: S0939‐6411(16)31020‐7.

Malik A (2016) Protein fusion tags for efficient expression and purification of recombinant proteins in the periplasmic space of E. coli. 3. Biotechnology 6: 44.

Malissard M, Zeng S and Berger EG (1999) The yeast expression system for recombinant glycosyltransferases. Glycoconjugate Journal 16: 125–139.

Mizutani K (2015) High‐throughput plasmid construction using homologous recombination in yeast: its mechanisms and application to protein production for X‐ray crystallography. Bioscience Biotechnology and Biochemistry 79: 1–10.

Retallack DM, Jin H and Chew L (2012) Reliable protein production in a Pseudomonas fluorescens expression system. Protein Expression and Purification 81: 157–165.

Scranton MA, Ostrand JT, Fields FJ and Mayfield SP (2015) Chlamydomonas as a model for biofuels and bio‐products production. Plant Journal 82: 523–531.

Structural Genomics Consortium, China Structural Genomics Consortium, Northeast Structural Genomics Consortium, et al. (2008) Protein production and purification. Nature Methods 5: 135–146.

Terpe K (2006) Overview of bacterial expression systems for heterologous protein production: from molecular and biochemical fundamentals to commercial systems. Applied Microbiology and Biotechnology 72: 211–222.

Vitale A and Pedrazzini E (2005) Recombinant pharmaceuticals from plants: the plant endomembrane system as bioreactor. Molecular Interventions 5: 216–225.

Welch M, Villalobos A, Gustafsson C and Minshull J (2011) Designing genes for successful protein expression. Methods in Enzymology 498: 43–66.

Yang H, Liu L and Xu F (2016) The promises and challenges of fusion constructs in protein biochemistry and enzymology. Applied Microbiology and Biotechnology 100: 8273–8281.

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

Further Reading

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

Ferrer‐Miralles N and Villaverde A (2013) Bacterial cell factories for recombinant protein production; expanding the catalogue. Microbial Cell Factories 12: 113.

Frutos R, Denise H, Vivares C, et al. (2008) Pharmaceutical proteins in plants. A strategic genetic engineering approach for the production of tuberculosis antigens. Annals of the New York Academy of Sciences 1149: 275–280.

Graumann K and Premstaller A (2006) Manufacturing of recombinant therapeutic proteins in microbial systems. Biotechnology Journal 1: 164–186.

Kost TA, Condreay JP and Jarvis DL (2005) Baculovirus as versatile vectors for protein expression in insect and mammalian cells. Nature Biotechnology 23: 567–575.

Nevalainen H and Peterson R (2014) Making recombinant proteins in filamentous fungi‐ are we expecting too much? Frontiers in Microbiology 5: 75.

Wang Y, Zhao S, Bai L, Fan J and Liu E (2013) Expression systems and species used for transgenic animal bioreactors. BioMed Research International 2013: 580463.

Yin J, Li G, Ren X and Herrler G (2007) Select what you need: a comparative evaluation of the advantages and limitations of frequently used expression systems for foreign genes. Journal of Biotechnology 127: 335–347.

Zerbs S, Frank AM and Collart FR (2009) Bacterial systems for production of heterologous proteins. Methods in Enzymology 463: 149–168.

Related Sub-Topics:

Protein Folding, Evolution and Degradation | Translation and Protein Synthesis | Basic Cell Biology, Protein, RNA and DNA | Sample Preparation in Structural Biology | Biosynthesis | Food and Industrial Microbiology | Techniques in Cell Biology | Techniques and Tools in Molecular Biology
Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

Article Title: Protein Production for Biotechnology
 
How to Cite close
Korajczyk, Peter J, Zerbs, Sarah, and Collart, Frank R(Oct 2017) Protein Production for Biotechnology. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0000983.pub3]