scholarly journals Epi‐CHO, an episomal expression system for recombinant protein production in CHO cells

2005 ◽  
Vol 91 (6) ◽  
pp. 670-677 ◽  
Author(s):  
Rajkumar Kunaparaju ◽  
Mimi Liao ◽  
Noelle‐Anne Sunstrom
Keyword(s):  
Recombinant Protein ◽  
Cho Cells ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Expression System

Construction of a binary transgenic gene expression system for recombinant protein production in the middle silk gland of the silkworm Bombyx mori

2009 ◽  
Vol 19 (3) ◽  
pp. 473-487 ◽  
Author(s):  
Ken-ichiro Tatematsu ◽  
Isao Kobayashi ◽  
Keiro Uchino ◽  
Hideki Sezutsu ◽  
Tetsuya Iizuka ◽  
...  
Keyword(s):  
Gene Expression ◽  
Recombinant Protein ◽  
Bombyx Mori ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Expression System ◽  
Silk Gland ◽  
Gene Expression System ◽  
Silkworm Bombyx Mori

scholarly journals Recombinant protein production provoked accumulation of ATP, fructose-1,6-bisphosphate and pyruvate in E. coli K12 strain TG1

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Jan Weber ◽  
Zhaopeng Li ◽  
Ursula Rinas
Keyword(s):  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Expression System ◽  
Limiting Factors ◽  
Expression Vectors ◽  
Metabolic Enzyme ◽  
E Coli ◽  
Metabolic Burden ◽  
Fructose 1,6 Bisphosphate

Abstract Background Recently it was shown that production of recombinant proteins in E. coli BL21(DE3) using pET based expression vectors leads to metabolic stress comparable to a carbon overfeeding response. Opposite to original expectations generation of energy as well as catabolic provision of precursor metabolites were excluded as limiting factors for growth and protein production. On the contrary, accumulation of ATP and precursor metabolites revealed their ample formation but insufficient withdrawal as a result of protein production mediated constraints in anabolic pathways. Thus, not limitation but excess of energy and precursor metabolites were identified as being connected to the protein production associated metabolic burden. Results Here we show that the protein production associated accumulation of energy and catabolic precursor metabolites is not unique to E. coli BL21(DE3) but also occurs in E. coli K12. Most notably, it was demonstrated that the IPTG-induced production of hFGF-2 using a tac-promoter based expression vector in the E. coli K12 strain TG1 was leading to persistent accumulation of key regulatory molecules such as ATP, fructose-1,6-bisphosphate and pyruvate. Conclusions Excessive energy generation, respectively, accumulation of ATP during recombinant protein production is not unique to the BL21(DE3)/T7 promoter based expression system but also observed in the E. coli K12 strain TG1 using another promoter/vector combination. These findings confirm that energy is not a limiting factor for recombinant protein production. Moreover, the data also show that an accelerated glycolytic pathway flux aggravates the protein production associated “metabolic burden”. Under conditions of compromised anabolic capacities cells are not able to reorganize their metabolic enzyme repertoire as required for reduced carbon processing.

Transcription factor engineering in CHO cells for recombinant protein production

2019 ◽  
Vol 39 (5) ◽  
pp. 665-679 ◽  
Author(s):  
Matías Gutiérrez-González ◽  
Yesenia Latorre ◽  
Roberto Zúñiga ◽  
Juan Carlos Aguillón ◽  
María Carmen Molina ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Recombinant Protein ◽  
Cho Cells ◽  
Recombinant Protein Production ◽  
Protein Production

scholarly journals The Goldilocks Approach: A Review of Employing Design of Experiments in Prokaryotic Recombinant Protein Production

2018 ◽  
Vol 5 (4) ◽  
pp. 89 ◽  
Author(s):  
Albert Uhoraningoga ◽  
Gemma Kinsella ◽  
Gary Henehan ◽  
Barry Ryan
Keyword(s):  
Recombinant Protein ◽  
Design Of Experiments ◽  
Prokaryotic Expression ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Expression System ◽  
Culture Conditions ◽  
Media Composition ◽  
High Yields ◽  
Soluble Recombinant Protein

The production of high yields of soluble recombinant protein is one of the main objectives of protein biotechnology. Several factors, such as expression system, vector, host, media composition and induction conditions can influence recombinant protein yield. Identifying the most important factors for optimum protein expression may involve significant investment of time and considerable cost. To address this problem, statistical models such as Design of Experiments (DoE) have been used to optimise recombinant protein production. This review examines the application of DoE in the production of recombinant proteins in prokaryotic expression systems with specific emphasis on media composition and culture conditions. The review examines the most commonly used DoE screening and optimisation designs. It provides examples of DoE applied to optimisation of media and culture conditions.

scholarly journals The PAR promoter expression system: modified lac promoters for controlled recombinant protein production in Escherichia coli

2021 ◽  
Author(s):  
Joanne Hothersall ◽  
Rita E. Godfrey ◽  
Christos Fanitsios ◽  
Tim W. Overton ◽  
Stephen J.W. Busby ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Expression System
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