Enhanced expression of soluble antibody fragments by low-temperature and overdosing with a nitrogen source
Abstract
Escherichia coli has been a primary host for the prokaryotic production of antibody fragments (Fabs) and has contributed to several successes in the pharmaceutical industry. Nevertheless, the requirement of disulfide bonds often results in low-yield fermentation and a lack of cost-effectiveness. Despite the improved production of functional Fabs by fermentation below 30 °C, the limited cellular growth needs further work. To address these issues, we investigated the effect of nitrogen supply on the cellular growth and the Fab productivity. We used the anti-human VEGF-A Fab as a model that exhibited poor expression at 37 °C regardless of the amount of nitrogen supplied during fermentation. In stark contrast, the expression yield of soluble Fab with a gross nitrogen supply of 6.91 g/L of broth throughout the fermentation at 25 °C was 332 mg/L. Furthermore, and increased nitrogen supply of 10.9 g/L significantly improved the yield of active form by 59.7% and the cellular growth rate by 39.3%. These results indicate that overdosing of a nitrogen source at low temperature is critical to Fab productivity in E. coli.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Several studies have demonstrated that yield of recombinant antibody fragments in E. coli significantly improves at growth temperature below 30 °C, likely due to reduction of translation rate that favors proper Ig-like folding and reduces aggregation.
“Several studies have demonstrated that the yield of recombinant antibody fragments in E. coli significantly improves at growth temperature below 30 °C, likely due to the reduction of translation rate that favors proper Ig-like folding and reduces aggregation [90,91,92].”
From Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments (2020) · cites this paper as [90]
Supplementing a nitrogen source at low temperature is critical for Fab productivity in E. coli fermentations.
“These results indicate that supplementing a nitrogen source at low temperature is critical for Fab productivity in E. coli fermentations [90].”
From Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments (2020) · cites this paper as [90]
Cites
Lowry, O. H., Carter, J., Ward, J. B., & Glaser, L. (1971) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)34144-4 · Cited by 350 · see citing works
Ukkonen, K., Veijola, J., Vasala, A., & Neubauer, P. (2013) · Microbial Cell Factories · doi:10.1186/1475-2859-12-73 · Cited by 65 · see citing works
Flagged first, then by citations
Cited by
Sorted by citations
Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments
Sandomenico, A., Sivaccumar, J. P., & Ruvo, M. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21176324 · Cited by 134 · see citing works
“While the preparation of whole antibody molecules requires eukaryotic expression systems, antibody fragments like Fabs, scFvs and other similar surrogates that lack the glycosylation, can be conveniently prepared in E. coli backgrounds.”
MethodInfluential
- Authors
- Kim, S. J., Ha, G. S., Lee, G., Lim, S. I., Lee, C. M., Yang, Y. H., Lee, J., Kim, J. E., Lee, J. H., Shin, Y., Kim, C.-W., & Lee, D. E.
- Affiliations
- CJ CheilJedang (South Korea) (KR)
- Korea University (KR)
- Corresponding author
- Kim, Chan-wha
- Lee, Dong Eok
- Venue
- Enzyme and Microbial Technology
- Publisher
- Elsevier BV
- Published
- 2018-08
- Volume
- 115
- Pages
- 9-15
- ISSN
- 0141-0229 (ISSN-L)
- OpenAlex
- W2795936909
- Cited by
- 17
- References
- 2
- Citations by year
- 2026: 2 · 2025: 3 · 2024: 1 · 2023: 2 · 2022: 2 · 2021: 2 · 2020: 2 · 2019: 3
- FWCI
- 0.62
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Enzyme and Microbial Technology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Keywords
- antibody fragments, recombinant protein expression, Escherichia coli fermentation, low-temperature fermentation, nitrogen source, disulfide bonds, expression yield
- MeSH terms
- Cold Temperature, Escherichia coli, Fermentation, Humans, Immunoglobulin Fragments, Nitrogen, Vascular Endothelial Growth Factor A
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
