Application of on-line OUR measurements to detect actions points to improve baculovirus-insect cell cultures in bioreactors

• Published in: Journal of biotechnology Vol. 125; no. 3; pp. 385 - 394
• Main Authors: Lecina, M., Soley, A., Gràcia, J., Espunya, E., Lázaro, B., Cairó, J.J., Gòdia, F.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 18-09-2006; Amsterdam Elsevier; New York, NY
• Subjects: Animals; Automation; Bacterial Infections - metabolism; Baculoviridae - isolation & purification; Baculovirus; Biological and medical sciences; Bioreactors - microbiology; Biotechnology; Biotechnology - methods; Cell Culture Techniques - methods; Fundamental and applied biological sciences. Psychology; Insect cells-baculovirus; Insecta - cytology; Insecta - metabolism; Insecta - microbiology; Insecta - physiology; Models, Theoretical; On-line monitoring; OUR; Oxygen - analysis; Oxygen - metabolism; Protein expression; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; Spodoptera - microbiology; Time Factors; Time of action; Time of harvesting; Time of infection; Viral infection; Viral Structural Proteins - genetics; Viral Structural Proteins - isolation & purification; Viral Structural Proteins - metabolism; Virus Cultivation - methods; Insect cells-baculovirus; Time of harvesting; Time of action; Protein expression; On-line monitoring; OUR; Viral infection; Time of infection; Cell culture; Insecta; Optimization; Protein; Virus; Infection; Bioreactor; Baculoviridae; Arthropoda; Invertebrata; Application; Monitoring
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2006.03.014

The continuous monitoring of a process based on the culture of Sf9 insect cells and infection by a baculovirus as a vector to obtain recombinant VP2 protein is studied. On-line OUR determination is based on the direct oxygen measurement in the cell culture vessel and the application of the dynamic method. This approximation allows a proper description of cell growth, with precise identification of the balanced cell growth end and the most important action times in the process, as virus infection time and final cell harvesting.
A detailed study of the OUR profiles allows on-line monitoring of the effects of infection and expression protein process, a tool enabling the automatisation of the protein production process in a baculovirus-insect cell system. These parameters have been defined as time of action (TOAs), and include the most relevant actions to take in these type of processes: time of infection (TOI), time of feeding (TOF) and time of harvesting (TOH).