Optimized signal peptides for the development of high expressing CHO cell lines

  title={Optimized signal peptides for the development of high expressing CHO cell lines},
  author={Lars Kober and Christoph Zehe and Juergen Bode},
  journal={Biotechnology and Bioengineering},
Recombinant biotherapeutic proteins such as monoclonal antibodies are mostly produced in Chinese hamster ovary (CHO) cells and pharmaceutical companies are interested in an appropriate platform technology for the development of large‐scale production processes. A major aim of our study was therefore to improve the secretion efficiency of a recombinant biotherapeutic antibody by optimizing signal peptides. Reporter molecules such as gaussia and vargula luciferase or secreted alkaline phosphatase… 

Efficient mAb production in CHO cells with optimized signal peptide, codon, and UTR

Antibody drugs have been used to treat a number of diseases successfully. Producing antibodies with high yield and quality is necessary for clinical applications of antibodies. For a candidate

Improved production and purification of recombinant proteins from mammalian expression systems

Preliminary data generated from transient expression in HEK293F cells could be generalised to predict that of proteinStability and effector function was maintained across transient and stable production methods at all scales, indicating that preliminary data generated in this work could begeneralised to predictions of protein stably expressed in CHO cell populations.

Development of an in vitro screening system for synthetic signal peptide in mammalian cell-based protein production

The overall results indicate the utility of a novel in vitro screening system for synthetic signal peptide for mammalian cell-based protein production and identify novel signal peptides that could increase Fc-fusion protein production in mammalian cells.

Factors Affecting the Expression of Recombinant Protein and Improvement Strategies in Chinese Hamster Ovary Cells

Here, the factors affecting the expression of recombinant protein and improvement strategies in CHO cells are reviewed and the inherent characteristics of molecular will also affect the production of protein.

Computational Design for the Production of Secretory Recombinant Codon-Optimized Human Stem Cell Factor (SCF) in Chinese Hamster Ovary (CHO) Cells With an Appropriate Signal Peptide: An Intensive In Silico Study

The use of this approach may, therefore, lead to the production of highly efficient recombinant hSCF, which would be feasible for the mass production of this factor for therapeutic purposes.

Overcoming the Refractory Expression of Secreted Recombinant Proteins in Mammalian Cells through Modification of the Signal Peptide and Adjacent Amino Acids

This study identifies the role of different N-terminal signal peptides and residues immediately downstream, in influencing the level of secreted recombinant protein obtained from suspension HEK293 cells, and describes a strategy that could enable recombinant proteins that have so far proved refractory to expression in HEK 293 cells, to be produced in sufficient quantities to answer important biological questions.

Expression of Recombinant Antibodies

This review focuses on current antibody production systems including their usability for different applications, and recent developments of glycosylation-engineered yeast, insect cell lines, and transgenic plants are promising to obtain antibodies with “human-like” post-translational modifications.

Evaluation of artificial signal peptides for secretion of two lysosomal enzymes in CHO cells.

The results indicate that altering signal peptide can modulate the secretion of recombinant lysosome enzymes and will enable lysOSomal enzyme production for clinical use.



Improving mammalian cell factories : The selection of signal peptide has a major impact on recombinant protein synthesis and secretion in mammalian cells

The results described here indicate the vital importance of selection of the signal peptide when aiming to produce maximal amounts of recombinant protein in a mammalian expression system.

Comparison of different signal peptides for protein secretion in nonlytic insect cell system.

Development of a novel ER stress based selection system for the isolation of highly productive clones

It is concluded, that the novel ER stress based selection system developed during this study is suitable to identify and isolate clones with a high level of antibody expression.

A study of monoclonal antibody‐producing CHO cell lines: What makes a stable high producer?

The herein described extensive characterization studies could help understand the limitations to high‐level, stable recombinant protein production and find ways to improving and accelerating the process for high‐producer cell line generation and selection.

Recombinant expression systems in the pharmaceutical industry

  • F. Schmidt
  • Biology
    Applied Microbiology and Biotechnology
  • 2004
To fully exploit the secretory capacity of fungal species, a deeper understanding of their posttranslational modification physiology will be necessary to steer the degree and pattern of glycosylation, which influences both folding and secretion efficiency.

Regulation of Recombinant Monoclonal Antibody Production in Chinese Hamster Ovary Cells: A Comparative Study of Gene Copy Number, mRNA Level, and Protein Expression

The study reveals that the MTX‐mediated MAb overexpression results from both an increase in the gene copy number and more efficient transcription of each gene copy, and the HC to LC gene copy ratio may affect the protein yield in MAb expression systems.

Production of recombinant protein therapeutics in cultivated mammalian cells

  • F. Wurm
  • Biology, Engineering
    Nature Biotechnology
  • 2004
Recently, the productivity of mammalian cells cultivated in bioreactors has reached the gram per liter range in a number of cases, a more than 100-fold yield improvement over titers seen for similar processes in the mid-1980s.

Toward more efficient protein expression

It is shown that gene optimization can also be used to improve the production of a complex macromolecule, namely an antibody, and suggests that significant improvements in product yields can be achieved by gene optimization, which may facilitate the processing and translation of gene transcripts.