Optimization of norovirus virus-like particle production inPichia pastorisusing a real-time near-infrared bioprocess monitor
| Autor: | Cameron L. Bardliving, Jonathon T. Olesberg, Jaime Tome-Amat, Kaylee J. Lanz, Mitchell H. Maloy, Mark A. Arnold, Carl A. Batt, Stephanie A. Parker |
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| Rok vydání: | 2016 |
| Předmět: |
Glycerol
0106 biological sciences 0301 basic medicine Time Factors Biology medicine.disease_cause 01 natural sciences Pichia Microbiology Pichia pastoris 03 medical and health sciences chemistry.chemical_compound Virus-like particle 010608 biotechnology medicine Particle Size Bioprocess Spectroscopy Near-Infrared Chromatography Methanol Norovirus biology.organism_classification 030104 developmental biology chemistry Yield (chemistry) Fermentation Biotechnology |
| Zdroj: | Biotechnology Progress. 32:518-526 |
| ISSN: | 8756-7938 |
| Popis: | 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) . © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:518-526, 2016. |
| Databáze: | OpenAIRE |
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