Enhanced expression of soluble antibody fragments by low-temperature and overdosing with a nitrogen source
| Autor: | Jae Hee Lee, Gyong Sik Ha, Kim Se Jun, Chung Min Lee, Yong Uk Shin, Lee Jaemin, Sung In Lim, Yoo Hee Yang, Ju Eun Kim, Chan wha Kim, Lee Gyung-Hwa, Dong Eok Lee |
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| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Vascular Endothelial Growth Factor A Nitrogen chemistry.chemical_element Bioengineering medicine.disease_cause 01 natural sciences Applied Microbiology and Biotechnology Biochemistry Antibody fragments 03 medical and health sciences 010608 biotechnology medicine Escherichia coli Humans Food science Nitrogen source Immunoglobulin Fragments Disulfide bond Cold Temperature 030104 developmental biology chemistry Yield (chemistry) Fermentation Biotechnology |
| Zdroj: | Enzyme and microbial technology. 115 |
| ISSN: | 1879-0909 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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