Establishment of a production process for a novel vaccine candidate against Lawsonia intracellularis

Electronic Journal of Biotechnology

J.R. Toledo¹  E. Ramos¹  R. Montesino¹  N. Gutiérrez¹  J. Acosta¹  T. Ramos²  S. Salazar²  O. Sánchez³  A. González³  C. Altamirano⁴ 

[1] Biotechnology and Biopharmaceuticals Laboratory, Pathophysiology Department, School of Biological Sciences, Universidad de Concepción, Victor Lamas 1290, P.O. Box 160C, Concepción, Chile;Department of Live Sciences and Agriculture, Universidad de las Fuerzas Armadas ESPE, Quito, Ecuador;Recombinant Biopharmaceuticals Laboratory, Pharmacology Department, School of Biological Sciences, Universidad de Concepción, Victor Lamas 1290, P.O. Box 160C, Concepción, Chile;School of Biochemical Engineering, Pontificia Universidad Católica de Valparaíso, Chile;

关键词: Recombinant vaccine;    Proliferative enteropathy;    Fed-batch culture;    Lawsonia intracellularis;    Immune response;   

DOI  :  

来源: DOAJ

【 摘 要 】

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, Gutiérrez N, Sánchez 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

【 授权许可】

Unknown