Related Experiment Video
Updated: Mar 24, 2026
12:28
Bacterial Inner-membrane Display for Screening a Library of Antibody Fragments
Published on: October 15, 2016
12.2K
Enhancing full-length antibody production by signal peptide engineering
Yizhou Zhou1, Peter Liu2, Yutian Gan3
1Department of Early Stage Cell Culture, Genentech Inc., 1 DNA way, South San Francisco, CA, 94080, USA. zyz1977627@gmail.com.
Microbial Cell Factories
|March 4, 2016
Summary
Optimizing signal peptide hydrophobicity enhances heavy chain secretion, a key step for improving full-length monoclonal antibody (mAb) production in E. coli periplasm. This strategy boosts mAb yields by addressing secretion limitations.
Area of Science:
- Biotechnology
- Molecular Biology
- Protein Engineering
Background:
- Escherichia coli periplasmic secretion is a viable method for heterologous protein production, including antibodies.
- Optimizing the translation initiation region (TIR) enhances protein production, but yields for complex proteins like monoclonal antibodies (mAbs) require further improvement.
Purpose of the Study:
- To identify limitations in full-length mAb accumulation in the E. coli periplasm.
- To enhance heavy chain secretion and improve overall mAb production yields.
Main Methods:
- Investigated various signal peptides for heavy chain secretion at controlled TIR strengths.
- Conducted mutagenesis studies to assess the impact of signal peptide hydrophobic core (H-region) hydrophobicity.
- Evaluated heavy chain secretion efficiency and periplasmic mAb accumulation.
Main Results:
- Inefficient heavy chain secretion was identified as the bottleneck for full-length mAb accumulation.
- The signal peptide of disulfide oxidoreductase (DsbA) demonstrated superior heavy chain secretion efficiency.
- Signal peptide H-region hydrophobicity critically influences heavy chain secretion and mAb accumulation; increased hydrophobicity enhances secretion.
Conclusions:
- Signal peptide hydrophobicity is crucial for heavy chain secretion under controlled translational strengths.
- Modulating signal peptide H-region hydrophobicity and TIR strength offers a strategy to improve heavy chain secretion and full-length mAb production in E. coli.
