Preparation of Very-High-Yield Recombinant Proteins Using Novel High-Cell-Density Bacterial Expression Methods

2010 ◽  
Vol 2010 (8) ◽  
pp. pdb.prot5475 ◽  
Author(s):  
Victoria Murray ◽  
Jianglei Chen ◽  
Yuefei Huang ◽  
Qianqian Li ◽  
Jianjun Wang
Keyword(s):  
Cell Density ◽  
Recombinant Proteins ◽  
High Cell Density ◽  
Bacterial Expression ◽  
High Yield ◽  
High Cell ◽  
Very High

scholarly journals A novel fed-batch based cultivation method provides high cell-density and improves yield of soluble recombinant proteins in shaken cultures

2010 ◽  
Vol 9 (1) ◽  
pp. 11 ◽  
Author(s):  
Mirja Krause ◽  
Kaisa Ukkonen ◽  
Tatu Haataja ◽  
Maria Ruottinen ◽  
Tuomo Glumoff ◽  
...  
Keyword(s):  
Cell Density ◽  
Recombinant Proteins ◽  
High Cell Density ◽  
High Cell ◽  
Fed Batch ◽  
Cultivation Method

scholarly journals Very high cell density perfusion of CHO cells anchored in a non-woven matrix-based bioreactor

2015 ◽  
Vol 213 ◽  
pp. 28-41 ◽  
Author(s):  
Ye Zhang ◽  
Per Stobbe ◽  
Christian Orrego Silvander ◽  
Véronique Chotteau
Keyword(s):  
Cell Density ◽  
Cho Cells ◽  
High Cell Density ◽  
High Cell ◽  
Very High

scholarly journals Antibody capture process based on magnetic beads from very high cell density suspension

2021 ◽  
Author(s):  
Nils A. Brechmann ◽  
Hubert Schwarz ◽  
Per‐Olov Eriksson ◽  
Kristofer Eriksson ◽  
Atefeh Shokri ◽  
...  
Keyword(s):  
Cell Density ◽  
Magnetic Beads ◽  
High Cell Density ◽  
High Cell ◽  
Capture Process ◽  
Antibody Capture ◽  
Very High

scholarly journals Media optimization for SHuffle T7 Escherichia coli expressing SUMO-Lispro proinsulin by response surface methodology

2022 ◽  
Vol 22 (1) ◽  
Author(s):  
Aida Bakhshi Khalilvand ◽  
Saeed Aminzadeh ◽  
Mohammad Hossein Sanati ◽  
Fereidoun Mahboudi
Keyword(s):  
Escherichia Coli ◽  
Cell Density ◽  
Shake Flask ◽  
High Cell Density ◽  
Fold Increase ◽  
Soluble Expression ◽  
High Yield ◽  
High Cell ◽  
E Coli ◽  
High Cell Density Fermentation

Abstract Background SHuffle is a suitable Escherichia coli (E. coli) strain for high yield cytoplasmic soluble expression of disulfide-bonded proteins such as Insulin due to its oxidative cytoplasmic condition and the ability to correct the arrangement of disulfide bonds. Lispro is an Insulin analog that is conventionally produced in E. coli as inclusion bodies (IBs) with prolonged production time and low recovery. Here in this study, we aimed to optimize cultivation media composition for high cell density fermentation of SHuffle T7 E. coli expressing soluble Lispro proinsulin fused to SUMO tag (SU-INS construct) to obtain high cell density fermentation. Results Factors including carbon and nitrogen sources, salts, metal ions, and pH were screened via Plackett–Burman design for their effectiveness on cell dry weight (CDW) as a measure of cell growth. The most significant variables of the screening experiment were Yeast extract and MgCl2 concentration, as well as pH. Succeedingly, The Central Composite Design was utilized to further evaluate and optimize the level of significant variables. The Optimized media (OM-I) enhanced biomass by 2.3 fold in the shake flask (2.5 g/L CDW) that reached 6.45 g/L (2.6 fold increase) when applied in batch culture fermentation. The efficacy of OM-I media for soluble expression was confirmed in both shake flask and fermentor. Conclusion The proposed media was suitable for high cell density fermentation of E. coli SHuffle T7 and was applicable for high yield soluble expression of Lispro proinsulin.

scholarly journals pH AND POPULATION DENSITY IN THE REGULATION OF ANIMAL CELL MULTIPLICATION

1971 ◽  
Vol 51 (3) ◽  
pp. 686-702 ◽  
Author(s):  
H. Rubin
Keyword(s):  
Cell Migration ◽  
Cell Density ◽  
Plasma Membranes ◽  
High Cell Density ◽  
Low Ph ◽  
Cell Multiplication ◽  
High Cell ◽  
High Ph ◽  
Cell Densities ◽  
Very High

Sparse and dense cultures of chick embryo cells were affected differently by pH. The rates of cell multiplication and of thymidine-3H incorporation into DNA of dense cultures were increased as the pH was increased from 6.6 to 7.6. At pH higher than 7.6 the rate of multiplication decreased slightly in the dense cultures, but the rate of thymidine-3H incorporation continued to increase. The discrepancy was due in part to cell death and detachment at very high pH, and in part to a more rapid uptake of thymidine-3H at very high pH. Sparse cultures were much less sensitive to pH reduction and, when a suitably conditioned medium was used to minimize cell damage, very sparse cultures grew almost as well at pH 6.7 as at higher pH. The rates of cell multiplication and thymidine-3H incorporation at low pH decreased in the initially sparse cultures before they reached confluent cell densities. There was no microscope evidence of direct contact between plasma membranes of cells at these densities although the parallel orientation indicated that the cells were influencing locally each other's behavior. Even at much higher cell densities, electron microscopy revealed large intercellular gaps partly filled with a fragmentary electron-opaque material suspected to be glycoprotein. Wounding experiments showed that pH affected cell migration in a manner similar to its effects on cell multiplication. Low pH inhibited cell migration, but those cells which migrated into the denuded region multiplied as rapidly at low pH as at high pH. The effects of pH on growth were correlated with effects on the uptake of 2-deoxyglucose-3H. Dense populations of cells inhibited by low pH were stimulated to incorporate thymidine-3H by the addition of small amounts of diethylaminoethyl-dextran. Rous sarcoma cells at high cell density were less sensitive to pH than were normal cells at the same density, but were more sensitive than sparse normal cultures. The results suggest that cell growth is inhibited through the combined effects of both lowered pH and high cell density on cell surface permeability.

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