期刊
JOURNAL OF BIOTECHNOLOGY
卷 102, 期 3, 页码 281-290出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/S0168-1656(03)00049-X
关键词
Pichia pastoris; fed batch; heterologous protein; methanol induction; viability; immunofluorescence
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.
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