High level production and purification of human interferon α2b in high cell density culture of Pichia pastoris

2008 ◽  
Vol 42 (2) ◽  
pp. 173-180 ◽  
Author(s):  
Atef Ayed ◽  
Imen Rabhi ◽  
Koussay Dellagi ◽  
Héla Kallel
Keyword(s):  
Pichia Pastoris ◽  
Cell Density ◽  
High Cell Density ◽  
Human Interferon ◽  
High Cell Density Culture ◽  
High Cell ◽  
High Level ◽  
Level Production ◽  
Interferon Α2b

scholarly journals The Presence of N-Terminal Secretion Signal Sequences Leads to Strong Stimulation of the Total Expression Levels of Three Tested Medically Important Proteins during High-Cell-Density Cultivations of Escherichia coli

2006 ◽  
Vol 73 (3) ◽  
pp. 906-912 ◽  
Author(s):  
H. Sletta ◽  
A. T�ndervik ◽  
S. Hakv�g ◽  
T. E. Vee Aune ◽  
A. Nedal ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Density ◽  
High Cell Density ◽  
Signal Sequence ◽  
High Cell ◽  
Signal Sequences ◽  
Expression Levels ◽  
Gm Csf ◽  
High Level ◽  
Level Production

ABSTRACT Genetic optimizations to achieve high-level production of three different proteins of medical importance for humans, granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon alpha 2b (IFN-α2b), and single-chain antibody variable fragment (scFv-phOx), were investigated during high-cell-density cultivations of Escherichia coli. All three proteins were poorly expressed when put under control of the strong Pm/xylS promoter/regulator system, but high volumetric yields of GM-CSF and scFv-phOx (up to 1.7 and 2.3 g/liter, respectively) were achieved when the respective genes were fused to a translocation signal sequence. The choice of signal sequence, pelB, ompA, or synthetic signal sequence CSP, displayed a high and specific impact on the total expression levels for these two proteins. Data obtained by quantitative PCR confirmed relatively high in vivo transcript levels without using a fused signal sequence, suggesting that the signal sequences mainly stimulate translation. IFN-α2b expression remained poor even when fused to a signal sequence, and an alternative IFN-α2b coding sequence that was optimized for effective expression in Escherichia coli was therefore synthesized. The total expression level of this optimized gene remained low, while high-level production (0.6 g/liter) was achieved when the gene was fused to a signal sequence. Together, our results demonstrate a critical role of signal sequences for achieving industrial level expression of three human proteins in E. coli under the conditions tested, and this effect has to our knowledge not previously been systematically investigated.

Recovery of recombinant β-glucosidase by expanded bed adsorption from Pichia pastoris high-cell-density culture broth

2006 ◽  
Vol 122 (1) ◽  
pp. 86-98 ◽  
Author(s):  
Theppanya Charoenrat ◽  
Mariena Ketudat-Cairns ◽  
Mehmedalija Jahic ◽  
Sven-Olof Enfors ◽  
Andres Veide
Keyword(s):  
Pichia Pastoris ◽  
Cell Density ◽  
High Cell Density ◽  
Culture Broth ◽  
Expanded Bed Adsorption ◽  
High Cell Density Culture ◽  
High Cell ◽  
Expanded Bed

Combined transcriptome and proteome analysis ofEscherichia coli during high cell density culture

2003 ◽  
Vol 81 (7) ◽  
pp. 753-767 ◽  
Author(s):  
Sung Ho Yoon ◽  
Mee-Jung Han ◽  
Sang Yup Lee ◽  
Ki Jun Jeong ◽  
Jong-Shin Yoo
Keyword(s):  
Cell Density ◽  
High Cell Density ◽  
Proteome Analysis ◽  
Ofescherichia Coli ◽  
High Cell Density Culture ◽  
High Cell
Sign in / Sign up

Export Citation Format

Share Document