University of Natural Resources and Life Sciences, Vienna (BOKU) - Research portal
Selected Publication:
Hohenblum, H; Borth, N; Mattanovich, D.
(2003):
Assessing viability and cell-associated product of recombinant protein producing Pichia pastoris with flow cytometry.
J Biotechnol. 2003; 102(3):281-290
FullText
FullText_BOKU
- Abstract:
- This paper describes the establishment of flow cytometric methods for recombinant Pichia pastoris strains, and their application to a lab scale fed batch fermentation. Using a strain which secretes human trypsinogen, the viability and the product which remained associated to the cell were measured with propidium iodide and immunofluorescent staining, respectively. Viability decreases significantly below 70% during the methanol fed batch phase, indicating a stress situation triggered by the fermentation conditions. Cell associated product is accumulated earlier after methanol induction than secreted product. These data demonstrate that flow cytometry is a powerful tool for the analysis and optimization of recombinant protein production processes, and they indicate the need to further improve a widely used fermentation protocol for P. pastoris. (C) 2003 Elsevier Science B.V. All rights reserved.
- Authors BOKU Wien:
- Borth Nicole
- Katinger Hermann
- Mattanovich Diethard
- Find related publications in this database (using NML MeSH Indexing)
- Bioreactors - microbiology
- Cell Culture Techniques - methods
- Cell Division - methods
- Cell Survival - methods
- Flow Cytometry - methods
- Humans - methods
- Methanol - metabolism
- Pichia - cytology
- Pilot Projects - cytology
- Recombinant Proteins - biosynthesis
- Trypsinogen - biosynthesis
- Find related publications in this database (Keywords)
- Pichia pastoris
- fed batch
- heterologous protein
- methanol induction
- viability
- immunofluorescence
Altmetric: