High Throughput Bioreactor Validation and Use in Mammalian Cell Culture Process Change Studies Utilizing Gene Expression Data

Title	High Throughput Bioreactor Validation and Use in Mammalian Cell Culture Process Change Studies Utilizing Gene Expression Data
Author	Hanson, Michael
Advisors	Moreira, Antonio; Rao, Govind
Program	Engineering, Chemical and Biochemical
UMBC Department	Chemical and Biochemical Engineering
Document Type	dissertation
Sponsors	University of Maryland, Baltimore County (UMBC)
Keywords	high throughput bioprocessing; optical sensor; disposable bioreactor; gene expression; DNA microarray; cell culture process changes; Engineering, Chemical (0542)
Date Issued	2008-02-05; *
Abstract	During upstream bioprocess development, a high producing production organism is selected, an appropriate growth media is formulated, and optimal bioreactor growth conditions and cultivation schemes are established. Hundreds of experiments must be carried out to accomplish these objectives. The first aspect of my dissertation research was to improve the tools available for carrying out this multitude of development experiments. First, optical sensors for dissolved oxygen (DO) and pH were compared to traditional electrochemical probes during hybridoma cell culture in a 5 L bioreactor. There was a 98.7 and 99.7% correlation between our DO and pH optical sensors and the electrochemical probes, respectively, throughout the duration of the experiment. These optical sensors were also employed in a high throughput bioreactor (HTBR) system capable of simultaneously carrying out twelve 35 ml cell culture experiments. We validated the HTBR for use in process change studies by testing bioreactor reproducibility in terms of sensor and cell culture data. We also showed that sensor instrumentation had no impact on cell physiology at the transcript level. Cell culture comparisons were carried out between the HTBR, a 5 L lab scale stirred-tank bioreactor, and a disposable bag bioreactor. Cells grew similarly in all three systems, but achieved a higher productivity in the bag bioreactor. In addition to during development, changes are made at the cell culture level throughout the biopharmaceutical product lifetime to allow for improvements such as enhanced expression systems, different media components, increased yields, scale-up, and new technologies. The second aspect of my dissertation research was to evaluate the utility of using DNA microarrays to screen cell culture process changes for their potential to adversely affect product quality. We studied the impact of sodium butyrate addition, increased osmolality, and change in bioreactor type. All process changes resulted in significant differences in gene expression and culture behavior. In summary, we have shown that DNA microarrays can be used to profile the transcriptional response of industrially relevant cell culture process changes.
Identifier	1136
Format	application:pdf
Language	en
Collection	UMBC Theses and Dissertations
Rights Statement	This item may be protected under Title 17 of the U.S. Copyright Law. It is made available by UMBC for non-commercial research and education. For permission to publish or reproduce, please see http://aok.lib.umbc.edu/specoll/repro.php or contact Special Collections at speccoll(at)umbc.edu.

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High Throughput Bioreactor Validation and Use in Mammalian Cell Culture Process Change Studies Utilizing Gene Expression Data

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