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

doi:10.1002/btpr.2224

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

Journal article

Peer reviewed

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 and Mark A Arnold

Biotechnology progress, Vol.32(2), pp.518-526

03/2016

DOI: 10.1002/btpr.2224

PMID: 26748930

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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) . © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:518-526, 2016.

Fermentation

Glycerol - analysis

Glycerol - metabolism

Methanol - analysis

Methanol - metabolism

Norovirus - chemistry

Norovirus - metabolism

Particle Size

Pichia - chemistry

Pichia - metabolism

Spectroscopy, Near-Infrared

Time Factors

Details

Title: Subtitle: Optimization of norovirus virus-like particle production in Pichia pastoris using a real-time near-infrared bioprocess monitor
Creators: Stephanie A Parker - Dept. of Biomedical Engineering, Cornell University, 357 Stocking Hall Ithaca, NY 14853, Ithaca, NY
Mitchell H Maloy - Dept. of Biological Engineering, Cornell University, Ithaca, NY
Jaime Tome-Amat - Dept. of Microbiology, Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, NY
Cameron L Bardliving - Dept. of Food Science, Cornell University, Ithaca, NY
Carl A Batt - Dept. of Food Science, Cornell University, Ithaca, NY
Kaylee J Lanz - ASL Analytical, Inc, 2500 Crosspark Rd, Coralville, IA
Jonathon T Olesberg - ASL Analytical, Inc, 2500 Crosspark Rd, Coralville, IA
Mark A Arnold - ASL Analytical, Inc, 2500 Crosspark Rd, Coralville, IA
Resource Type: Journal article
Publication Details: Biotechnology progress, Vol.32(2), pp.518-526
DOI: 10.1002/btpr.2224
PMID: 26748930
ISSN: 8756-7938
eISSN: 1520-6033
Grant note: name: Cornell Center for Materials Research Shared Facilities; name: NSF MRSEC program, award: DMR-1120296
Language: English
Date published: 03/2016
Academic Unit: Physics and Astronomy; Center for Biocatalysis and Bioprocessing; Fraternal Order of Eagles Diabetes Research Center; Chemistry
Record Identifier: 9984216671502771

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