Listed below are a number of articles that provide a broad overview about using different expression systems for the production of recombinant proteins. Jump to each section by clicking the link below:

Bacteria - Yeast - Insect-cell/BEVS - Mammalian-cell - In Vitro

Bacterial expression system

Papaneophytou, C.P. et al. Statistical approaches to maximize recombinant protein expression in Escherichia coli: A general review. Protein Expression and Purification 94, 22–32 (2014) Link

Kudla, G. Coding-Sequence Determinants of Gene Expression in Escherichia coli. Science 324, 255-258 (2014) Link

Muntari, B., et al. Recombinant bromelain production in Escherichia coli: process optimization in shake flask culture by response surface methodology. AMB Express 2:12 (2012) Link

Rouet, R., et al. Expression of high-affinity human antibody fragments in bacteria. Nature Protocols 364VOL.7 NO.2 (2012) Link

Vincentelli, R. et al. High-throughput protein expression screening and purification in Escherichia coli, Methods 55, 65-72 (2011) Link

Unger, T., et al. Applications of the Restriction Free (RF) cloning procedure for molecular manipulations and protein expression. Journal of Structural Biology 172, 34-44 (2010). Link

Schein, C.H. Soluble Protein Expression in Bacteria. Encyclopedia of Industrial Biotechnology: Bioprocess, Bioseparation, and Cell Technology 1-20 (2010).

Francis, D.M. et al. Strategies to Optimize Protein Expression in E. coli. Current protocols in protein science Chapter 5, Unit 5 24 (2010). Link

Brondyk, W.H. Selecting an Appropriate Method for Expressing a Recombinant Protein. Guide to Protein Purification, Second Edition 463, 131-147 (2009). Link

Rosano G.L. et al. Rare codon content affects the solubility of recombinant proteins in a codon bias-adjusted Escherichia coli strain. Microbial Cell Factories 8:41 (2009) Link

Selleck, W. et al. Recombinant protein complex expression in E. coli. Current protocols in protein science Chapter 5, Unit 5 21 (2008). Link

Komar A.A A pause for thought along the co-translational folding pathway. Trends in Biochemical Sciences Vol.34 No.1 (2008) Link

Berrow, N.S. et al. A versatile ligation-independent cloning method suitable for high-throughput expression screening applications. Nucleic Acids Research 35, e45 (2007). Link

Cabrita, L.D., et al. A family of E. coli expression vectors for laboratory scale and high throughput soluble protein production. BMC Biotechnology 6, 12 (2006). Link

Duetz, W.A., et al. Methods for Intense Aeration, Growth, Storage, and Replication of Bacterial Strains in Microtiter Plates. Applied and Environmental Microbiology, Vol 66, No. 6, 2641–2646 (2006) Link

Waugh, D. Making the most of affinity tags. TRENDS in Biotechnology Vol.23 No.6 (2005) Link

Goulding C.W. et al. Protein production in Escherichia coli for structural studies by X-ray crystallography. Journal of Structural Biology 142 133–143 (2003) Link


Yeast expression system

Spadiut, O. et al. Microbials for the production of monoclonal antibodies and antibody fragments. Trends in Biotechnology 1:54-60 (2014) Link

Vogl, T. et al. New opportunities by synthetic biology for biopharmaceutical production in Pichia pastoris. Current Opinion in Biotechnology 6:1094-101 (2013) Link

Jiang Y. et al. Purification process development of a recombinant monoclonal antibody expressed in glycoengineered Pichia pastoris. Protein Expression and Purification 76 7–14 (2011) Link

Abad, S. et al. Stepwise engineering of a Pichia pastoris D-amino acid oxidase whole cell catalyst. Microbial Cell Factories, 9:24 (2010) Link

Bollok, M., Resina, D., Valero, F. & Ferrer, P. Recent patents on the Pichia pastoris expression system: expanding the toolbox for recombinant protein production. Recent Patents on Biotechnology 3, 192-201 (2009). Link

Cregg, J.M. et al. Expression in the yeast Pichia pastoris. Methods in Enzymology 463, 169-189 (2009). Link

Gong B. et al. Characterization of N-Linked Glycosylation on Recombinant Glycoproteins Produced in Pichia pastoris Using ESI-MS and MALDI-TOF. Methods in Molecular Biology, Glycomics: Methods and Protocols, vol. 534 Chapter 16 (2009) Link

Ghosalkar A. et al. Optimization of chemically defined medium for recombinant Pichia pastoris for biomass production. Bioresource Technology 99 7906–7910 (2008) Link

Hamilton, S.R. & Gerngross, T.U. Glycosylation engineering in yeast: the advent of fully humanized yeast. Current Opinion in Biotechnology 18, 387-392 (2007). Link

Lin-Cereghino, J., et al., Condensed protocol for competent cell preparation and transformation of the methylotrophic yeast Pichia pastoris. Biotechniques 38(1): 44–48. (2005)  Link

Suga M. et al. Cryopreservation of competent intact yeast cells for efficient electroporation. Yeast 16: 889-896. (2000) Link


Insect cell / BEVS expression system

Fernandes, F. et al. Insect cells as a production platform of complex virus-like particles Expert Rev. Vaccines12(2), 225–236 (2013) Link

Kost, T. Baculovirus Gene Delivery: A Flexible Assay Development Tool. Current Gene Therapy 10 168-173 (2010) Link

Airenne, K.J., et. al. In Vivo  Application and Tracking of Baculovirus. Current Gene Therapy, 2010 Link

Madhan, S. et al. Baculovirus as Vaccine Vectors. Current Gene Therapy 10 201-213 (2010) Link

Trowitzsch, S., et al.  New baculovirus expression tools for recombinant protein complex production. Journal of Structural Biology 172, 45-54 (2010). Link

Aucoin, M.G., et al.  Bioprocessing of baculovirus vectors: a review. Current Gene Therapy 10, 174-186 (2010). Link

Jarvis, D.L. Baculovirus-insect cell expression systems. Methods in Enzymology 463, 191-222 (2009). Link

Chung Y.C., et al., Expression, purification and characterization of enterovirus-71 virus-like particles. World J Gastroenterol; 12(6): 921-927 (2006) Link

Kost, T.A., et al.  Baculovirus as versatile vectors for protein expression in insect and mammalian cells. Nature Biotechnology 23, 567-575 (2005). Link

Weber, W. Optimisation of protein expression and establishment of the Wave Bioreactor for Baculovirus/insect cell culture. Cytotechnology 38: 77–85, (2002) Link


Mammalian cell expression system

Almo S.C., et al. Better and faster: improvements and optimization for mammalian recombinant protein production. Current Opinion in  Structural Biology. 26:39-43 (2014) Link

Bandaranayake A.D. et al. Recent advances in mammalian protein production. FEBS Letters. 588 2 253-60 (2014) Link

Zhang, J. Mammalian Cell Culture for Biopharmaceutical Production. Fermentation and Cell Culture Chapter 12 157-178 (2012) Link

Baldi. L., et al. Large-scale transfection of mammalian cells. Methods in Molecular Biology Volume 801 13-26 (2012) Link

Rodrigues, M.E. Technological Progresses in Monoclonal Antibody Production Systems. Biotechnol. Prog. Vol. 26, No. 2 (2010) Link

Hacker D.L. 25 years of recombinant proteins from reactor-grown cells—Where do we go from here? Biotechnology Advances 27 1023–1027 (2009) Link

Nettleship, J. et al., The Production of Glycoproteins by Transient Expression in Mammalian Cells. Methods in Molecular Biology: High Throughput Protein Expression and Purification, vol. 498 Chapter 16 (2009) Link

Lackner, A. A bicistronic baculovirus vector for transient and stable protein expression in mammalian cells. Analytical Biochemistry 380 146–148 (2008) Link

Matasci, M. et al. Recombinant therapeutic protein production in cultivated mammalian cells:current status and future prospects. Drug Discovery Today: Technologies 5, e37-e42 (2008). Link

Baldi, L., et al. Recombinant protein production by large-scale transient gene expression in mammalian cells: state of the art and future perspectives. Biotechnology Letters 29, 677-684 (2007). Link

Barnes, L.M., et al. Mammalian cell factories for efficient and stable protein expression. Current Opinion in Biotechnology 17 381–386 (2006) Link

Birch, J.R. et al. Antibody production. Advanced Drug Delivery Reviews 58 671 – 685 (2006) Link

Kunaparaju, R., et al., Epi-CHO, an Episomal  Expression System for Recombinant Protein Production in CHO Cells. Wiley Periodicals, (2005) Link

Wurm, F.M. Production of recombinant protein therapeutics in cultivated mammalian cells. Nature Biotechnology 22 No. 11 (2004) Link


In Vitro expression system

Johnston W & Alexandrov K (2014) Production of Eukaryotic Cell-Free Lysate from Leishmania tarentolae. Cell-Free Protein Synthesis, Methods in Molecular Biology, eds Alexandrov K & Johnston WA (Humana Press), Vol 1118, pp 1-15.

Kovtun O, Mureev S, Johnston W, & Alexandrov K (2010) Towards the construction of expressed proteomes using a Leishmania tarentolae based cell-free expression system. PLoS One 5(12):e14388.

Kovtun O, et al. (2011) Leishmania cell-free protein expression system. Methods 55(1):58-64.

Mureev S, Kovtun O, Nguyen UT, & Alexandrov K (2009) Species-independent translational leaders facilitate cell-free expression. Nat Biotechnol 27(8):747-752.