1. Effect of temperature on the production of a recombinant antivenom in fed-batch mode
- Author
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Balkiss Bouhaouala-Zahar, César Arturo Aceves Lara, Hazar Ghezal-Kraïem, Susana María Alonso Villela, Carine Bideaux, Luc Fillaudeau, Toulouse Biotechnology Institute (TBI), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Université de Tunis El Manar (UTM), Consejo Nacional de Ciencia y Tecnologia (CONACyT) (461347), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire des Venins et Biomolécules Thérapeutiques - Laboratory of Venoms and Therapeutic Biomolecules (LR11IPT08), Institut Pasteur de Tunis, and Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)
- Subjects
[SDV.BIO]Life Sciences [q-bio]/Biotechnology ,Kinetic modeling ,Antivenom ,Kinetics ,MESH: Batch Cell Culture Techniques ,medicine.disease_cause ,Applied Microbiology and Biotechnology ,law.invention ,MESH: Recombinant Proteins ,MESH: Bioreactors ,03 medical and health sciences ,Bioreactors ,law ,Fed-batch strategy ,medicine ,Bioreactor ,Escherichia coli ,MESH: Antivenins ,Food science ,Bioprocess ,030304 developmental biology ,Recombinant antivenom ,0303 health sciences ,Strain (chemistry) ,MESH: Escherichia coli ,Antivenins ,030306 microbiology ,Chemistry ,Induction temperature ,Temperature ,General Medicine ,MESH: Temperature ,Recombinant Proteins ,Batch Cell Culture Techniques ,Batch processing ,Recombinant DNA ,Nanobody ,Biotechnology - Abstract
In the pharmaceutical industry, nanobodies show promising properties for its application in serotherapy targeting the highly diffusible scorpion toxins. The production of recombinant nanobodies in Escherichia coli has been widely studied in shake flask cultures in rich medium. However, there are no upstream bioprocess studies of nanobody production in defined minimal medium and the effect of the induction temperature on the production kinetics. In this work, the effect of the temperature during the expression of the chimeric bispecific nanobody CH10-12 form, showing high scorpion antivenom potential, was studied in bioreactor cultures of E. coli. High biomass concentrations (25 g cdw/L) were achieved in fed-batch mode, and the expression of the CH10-12 nanobody was induced at temperatures 28, 29, 30, 33, and 37°C with a constant glucose feed. For the bispecific form NbF12-10, the induction was performed at 29°C. Biomass and carbon dioxide yields were reported for each culture phase, and the maintenance coefficient was obtained for each strain. Nanobody production in the CH10-12 strain was higher at low temperatures (lower than 30°C) and declined with the increase of the temperature. At 29°C, the CH10-12, NbF12-10, and WK6 strains were compared. Strains CH10-12 and NbF12-10 had a productivity of 0.052 and 0.021 mg/L/h of nanobody, respectively, after 13 h of induction. The specific productivity of the nanobodies was modeled as a function of the induction temperature and the specific growth rates. Experimental results confirm that low temperatures increase the productivity of the nanobody.Key points• Nanobodies with scorpion antivenom activity produced using two recombinant strains.• Nanobodies production was achieved in fed-batch cultures at different induction temperatures.• Low induction temperatures result in high volumetric productivities of the nanobody CH10-12.
- Published
- 2021