Establishment of a production process for a novel vaccine candidate against Lawsonia intracellularis
| Autor: | Jannel Acosta, Eduardo Ramos, Raquel Montesino, Jorge R. Toledo, Thelvia Ramos, Nicolas A. Gutiérrez, Santiago Salazar, Alaín González, Oliberto Sánchez, Claudia Altamirano |
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| Rok vydání: | 2021 |
| Předmět: |
0106 biological sciences
0301 basic medicine lcsh:Biotechnology Recombinant vaccine Biology 01 natural sciences Applied Microbiology and Biotechnology Proliferative enteropathy Lawsonia intracellularis Microbiology 03 medical and health sciences Antigen Immunity lcsh:TP248.13-248.65 010608 biotechnology Immune response Fed-batch culture lcsh:QH301-705.5 Lawsonia Bacteria Immunogenicity Vaccination Bacterial vaccine 030104 developmental biology lcsh:Biology (General) Biotechnology |
| Zdroj: | Electronic Journal of Biotechnology, Vol 50, Iss, Pp 45-52 (2021) |
| ISSN: | 0717-3458 |
| DOI: | 10.1016/j.ejbt.2021.01.002 |
| Popis: | Background Lawsonia intracellularis remains a problem for the swine industry worldwide. Previously, we designed and obtained a vaccine candidate against this pathogen based on the chimeric proteins: OMP1c, OMP2c, and INVASc. These proteins formed inclusion bodies when expressed in E. coli, which induced humoral and cellular immune responses in vaccinated pigs. Also, protection was demonstrated after the challenge. In this study, we established a production process to increase the yields of the three antigens as a vaccine candidate. Results Batch and fed-batch fermentations were evaluated in different culture conditions using a 2 L bioreactor. A fed-batch culture with a modified Terrific broth medium containing glucose instead of glycerol, and induced with 0.75 mM IPTG at 8 h of culture (11 g/L of biomass) raised the volumetric yield to 627.1 mg/L. Under these culture conditions, plasmid-bearing cells increased by 10% at the induction time. High efficiency in cell disruption was obtained at passage six using a high-pressure homogenizer and a bead mill. The total antigen recovery was 64% (400 mg/L), with a purity degree of 70%. The antigens retained their immunogenicity in pigs, inducing high antibody titers. Conclusions Considering that the antigen production process allowed an increment of more than 70-fold, this methodology constitutes a crucial step in the production of this vaccine candidate against L. intracellularis. How to cite: Salazar S, Gutierrez N, Sanchez O, et al. Establishment of a production process for a novel vaccine candidate against Lawsonia intracellularis. Electron J Biotechnol 2021. https://doi.org/10.1016/j.ejbt.2021.01.002 |
| Databáze: | OpenAIRE |
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