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Title of Journal: Mol Biotechnol

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Abbravation: Molecular Biotechnology

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Humana Press Inc

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DOI

10.1007/bf00716357

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ISSN

1559-0305

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Enhanced Production of Human Recombinant Proteins

Authors: T Tharmalingam K Sunley M Spearman M Butler
Publish Date: 2011/04/07
Volume: 49, Issue: 3, Pages: 263-276
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Abstract

Macroporous microcarriers entrap cells in a mesh network allowing growth to high densities and protect them from high shear forces in stirred bioreactor cultures We report the growth of Chinese hamster ovary CHO cells producing either recombinant human betainterferon βIFN or recombinant human tissueplasminogen activator tPA in suspension or embedded in macroporous microcarriers Cytopore 1 or 2 The microcarriers enhanced the volumetric production of both βIFN and tPA by up to 25 fold compared to equivalent suspension cultures of CHO cells Under each condition the cell specific productivity Q P was determined as units of product/cell per day based upon immunological assays Cells grown in Cytopore 1 microcarriers showed an increase in Q P with increasing cell densities up to a threshold of 1 × 108 cells/ml At this point the specific productivity was 25 fold higher than equivalent cells grown in suspension but cell densities above this threshold did not enhance Q P any further A positive linear correlation r 2 = 093 was determined between the specific productivity of each recombinant protein and the corresponding cell density for CHO cells grown in Cytopore 2 cultures With a cell density range of 25 × 106 to 3 × 108 cells/ml within the microcarriers there was a proportional increase in the specific productivity The highest specific productivity measured from the microcarrier cultures was ×5 that of suspension cultures The relationship between specific productivity and cell density within the microcarriers leads to higher yields of recombinant proteins in this culture system This could be attributed to the environment within the microcarrier matrix that may influence the state of cells that could affect protein synthesis or secretion


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Other Papers In This Journal:

  1. Production and Characterization of a Single-Chain Fv Antibody–Alkaline Phosphatase Fusion Protein Specific for Clenbuterol
  2. Improvement of Cephalosporin C Production by Recombinant DNA Integration in Acremonium chrysogenum
  3. Codon Optimization of the “Bos Taurus Chymosin” Gene for the Production of Recombinant Chymosin in Pichia pastoris
  4. Heterologous Overexpression, Purification and Characterisation of an Alcohol Dehydrogenase (ADH2) from Halobacterium sp. NRC-1
  5. Enhanced Extracellular Production of Heterologous Proteins in Bacillus subtilis by Deleting the C-terminal Region of the SecA Secretory Machinery
  6. Molecular Cloning and Characterization of a Malic Enzyme Gene from the Oleaginous Yeast Lipomyces starkeyi
  7. Hydrophobic Substitution of Surface Residues Affects Lipase Stability in Organic Solvents
  8. Identification of Suitable Endogenous Normalizers for qRT-PCR Analysis of Plasma microRNA Expression in Essential Hypertension
  9. Biosynthesis of Resveratrol in Blastospore of the Macrofungus Tremella fuciformis
  10. Heterologous Expression and Characterization of an Alcohol Dehydrogenase from the Archeon Thermoplasma acidophilum
  11. Plasma Antioxidants and Human Aging: A Study on Healthy Elderly Tunisian Population
  12. Importance of the residue Asp 290 on chain length selectivity and catalytic efficiency of recombinant Staphylococcus simulans lipase expressed in E. coli
  13. Fahd Al-Mulla (ed). Formalin-Fixed Paraffin-Embedded Tissue. Methods and Protocols. Methods in Molecular Biology 724. The Humana Press, ISBN 978-1-61779-054-6
  14. The Peptide Derived from erbB2 Auto-Inhibitor Herstatin Shared in the Same Epitope and Function with Functional Antibody 2C4
  15. End Joining-Mediated Gene Expression in Mammalian Cells Using PCR-Amplified DNA Constructs that Contain Terminator in Front of Promoter
  16. Identification of Functional Regions in the Rhodospirillum rubrum l -Asparaginase by Site-Directed Mutagenesis
  17. Expression of an Acid Urease with Urethanase Activity in E. coli and Analysis of Urease Gene

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