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Rezaie F, Davami F, Mansouri K, et al. Cytosolic expression of functional Fab fragments in Escherichia coli using a novel combination of dual SUMO expression cassette and EnBase. J Appl Microbiol. 2017;123(1):134-144doi: 10.1111/jam.13483.
Rezaie, F., Davami, F., Mansouri, K., Agha Amiri, S., Fazel, R., Mahdian, R., Davoudi, N., Enayati, S., Azizi, M., & Khalaj, V. (2017). Cytosolic expression of functional Fab fragments in Escherichia coli using a novel combination of dual SUMO expression cassette and EnBase. Journal of applied microbiology, 123(1), 134-144. https://doi.org/10.1111/jam.13483
Rezaie, F, et al. "Cytosolic expression of functional Fab fragments in Escherichia coli using a novel combination of dual SUMO expression cassette and EnBase." Journal of applied microbiology vol. 123,1 (2017): 134-144. doi: https://doi.org/10.1111/jam.13483
Rezaie F, Davami F, Mansouri K, Agha Amiri S, Fazel R, Mahdian R, Davoudi N, Enayati S, Azizi M, Khalaj V. Cytosolic expression of functional Fab fragments in Escherichia coli using a novel combination of dual SUMO expression cassette and EnBase. J Appl Microbiol. 2017 Jul;123(1):134-144. doi: 10.1111/jam.13483. Epub 2017 Jun 09. PMID: 28482126.
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J Appl Microbiol. 2017 Jul;123(1):134-144. doi: 10.1111/jam.13483. Epub 2017 Jun 09.

Cytosolic expression of functional Fab fragments in Escherichia coli using a novel combination of dual SUMO expression cassette and EnBase.

Journal of applied microbiology

F Rezaie, F Davami, K Mansouri, S Agha Amiri, R Fazel, R Mahdian, N Davoudi, S Enayati, M Azizi, V Khalaj

Affiliations

  1. Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.
  2. Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran.
  3. Molecular Medicine Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran.

PMID: 28482126 DOI: 10.1111/jam.13483

Abstract

AIMS: The Escherichia coli expression system is highly effective in producing recombinant proteins. However, there are some limitations in this system, especially in obtaining correctly folded forms of some complex proteins such as Fab fragments. To improve the solubility and folding quality of Fab fragments, we have examined the effect of simultaneous application of a SUMO fusion tag, EnBase

METHODS AND RESULTS: A bicistronic gene construct was designed to express an antivascular endothelial growth factor (VEGF) Fab fragment as a model system. The construct contained a dual SUMO fusion gene fragment to encode SUMO-tagged heavy and light chains. While the expression of the construct in batch cultures of BL21 or SHuffle

CONCLUSIONS: This study demonstrated that the combination of SUMO fusion technology, EnBase

SIGNIFICANCE AND THE IMPACT OF THE STUDY: The presented strategy provides not only a novel method to produce soluble and active form of an anti-VEGF Fab but also may use in the efficient production of other antibody fragments.

© 2017 The Society for Applied Microbiology.

Keywords: E. coli expression system; E. coli redox mutant; Fed-batch culture; cytosolic expression; functional fab fragment; recombinant antibody

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