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Establishment of a production process for a novel vaccine candidate against Lawsonia intracellularis
Salazar, S; Gutiérrez, N; Sánchez, O; Ramos, E; González, A; Acosta, J; Ramos, T; Altamirano, C; Toledo, JR; Montesino, R.
  • Salazar, S; Universidad de las Fuerzas Armadas ESPE. Department of Live Sciences and Agriculture. Quito. EC
  • Gutiérrez, N; Universidad de Concepción. School of Biological Sciences. Pathophysiology Department. Concepción. CL
  • Sánchez, O; Universidad de Concepción. School of Biological Sciences. Pharmacology Department. Concepción. CL
  • Ramos, E; Universidad de Concepción. School of Biological Sciences. Pathophysiology Department. Concepción. CL
  • González, A; Universidad de Concepción. School of Biological Sciences. Pharmacology Department. Concepción. CL
  • Acosta, J; Universidad de Concepción. School of Biological Sciences. Pathophysiology Department. Concepción. CL
  • Ramos, T; Universidad de las Fuerzas Armadas ESPE. Department of Live Sciences and Agriculture. Quito. EC
  • Altamirano, C; Pontificia Universidad Católica de Valparaíso. School of Biochemical Engineering. CL
  • Toledo, JR; Universidad de Concepción. School of Biological Sciences. Pathophysiology Department. Concepción. CL
  • Montesino, R; Universidad de Concepción. School of Biological Sciences. Pathophysiology Department. Concepción. CL
Electron. j. biotechnol ; 50: 45-52, Mar. 2021. tab, graf
Article in English | LILACS | ID: biblio-1292328
ABSTRACT

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.
Subject(s)

Full text: Available Index: LILACS (Americas) Main subject: Swine Diseases / Bacterial Vaccines / Lawsonia Bacteria / Desulfovibrionaceae Infections Limits: Animals Language: English Journal: Electron. j. biotechnol Journal subject: Biotechnology Year: 2021 Type: Article Affiliation country: Chile / Ecuador Institution/Affiliation country: Pontificia Universidad Católica de Valparaíso/CL / Universidad de Concepción/CL / Universidad de las Fuerzas Armadas ESPE/EC

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Full text: Available Index: LILACS (Americas) Main subject: Swine Diseases / Bacterial Vaccines / Lawsonia Bacteria / Desulfovibrionaceae Infections Limits: Animals Language: English Journal: Electron. j. biotechnol Journal subject: Biotechnology Year: 2021 Type: Article Affiliation country: Chile / Ecuador Institution/Affiliation country: Pontificia Universidad Católica de Valparaíso/CL / Universidad de Concepción/CL / Universidad de las Fuerzas Armadas ESPE/EC