Substrate-source flexibility of an exponential-fed perfusion process to produce plasmid DNA for use as leishmaniasis vaccine

Aurora García-Rendón, Angelica García-Rendón, Roberto Z Guzman, Armando Tejeda-Mansir

Research output: Contribution to journalArticle

Abstract

The use of plasmid DNA (pDNA) for human vaccines is a novel approach against leishmaniasis, a neglected tropical disease with severe clinical manifestations. The development of feasible bioprocesses to obtain such vaccines is a public-health priority. The aim of this work was to investigate the substrate-source flexibility of an exponential-fed perfusion (EFP) system to produce the plasmid pVAX1-NH36 for use as a leishmaniasis vaccine. Batch and EFP cultures were conducted using Escherichia coli DH5α as a host and glucose or glycerol as a carbon source. The culture kinetics of the cell, substrate and plasmid concentrations were measured. Mathematical kinetics models were fitted to experimental data and used to describe the system comportment (r2 > 0.95). Plasmid productivities of 13.3 mg/(L h) using glucose and 19.4 mg/(L h) using glycerol were obtained. These levels represent a 1–3-fold increase in performance index compared with previously reported cultures using E. coli DH5α. The novel aspect of this work is the demonstration of the flexibility of EFP cultures for production of pDNA vaccines. Our data suggest that E. coli engineering to increase pDNA production using glucose can be circumvented with an EFP culture, reducing the host strain development costs. In addition, the greater productivity of EFP cultures entails a reduction in manufacturing costs.

Original languageEnglish (US)
JournalBiotechnology and Biotechnological Equipment
DOIs
StateAccepted/In press - Jan 1 2018

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Leishmaniasis Vaccines
Plasmids
Perfusion
DNA
DNA Vaccines
Escherichia coli
Glucose
Glycerol
Neglected Diseases
Costs and Cost Analysis
Health Priorities
Leishmaniasis
Theoretical Models
Carbon
Vaccines
Public Health
Cell Culture Techniques

Keywords

  • Growth modelling
  • perfusion culture
  • plasmid DNA
  • process flexibility
  • vaccine

ASJC Scopus subject areas

  • Biotechnology

Cite this

Substrate-source flexibility of an exponential-fed perfusion process to produce plasmid DNA for use as leishmaniasis vaccine. / García-Rendón, Aurora; García-Rendón, Angelica; Guzman, Roberto Z; Tejeda-Mansir, Armando.

In: Biotechnology and Biotechnological Equipment, 01.01.2018.

Research output: Contribution to journalArticle

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