Optimization of norovirus virus-like particle production in Pichia pastoris using a real-time near-infrared bioprocess monitor

Stephanie A. Parker, Mitchell H. Maloy, Jaime Tome-Amat, Cameron L. Bardliving, Carl A. Batt, Kaylee J. Lanz, Jonathon T. Olesberg, Mark A. Arnold

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

The production of norovirus virus-like particles (NoV VLPs) displaying NY-ESO-1 cancer testis antigen in Pichia pastoris BG11 Mut+ has been enhanced through feed-strategy optimization using a near-infrared bioprocess monitor (RTBio® Bioprocess Monitor, ASL Analytical, Inc.), capable of monitoring and controlling the concentrations of glycerol and methanol in real-time. The production of NoV VLPs displaying NY-ESO-1 in P. pastoris has potential as a novel cancer vaccine platform. Optimization of the growth conditions resulted in an almost two-fold increase in the expression levels in the fermentation supernatant of P. pastoris as compared to the starting conditions. We investigated the effect of methanol concentration, batch phase time, and batch to induction transition on NoV VLP-NY-ESO-1 production. The optimized process included a glycerol transition phase during the first 2 h of induction and a methanol concentration set point of 4 g L-1 during induction. Utilizing the bioprocess monitor to control the glycerol and methanol concentrations during induction resulted in a maximum NoV VP1-NY-ESO-1 yield of 0.85 g L-1.

Original languageEnglish
Pages (from-to)518-526
Number of pages9
JournalBiotechnology Progress
Volume32
Issue number2
DOIs
StatePublished - 1 Mar 2016

Keywords

  • Bioprocess monitor
  • Fed-batch fermentation
  • NY-ESO-1
  • Norovirus virus-like particles

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