An essential part of the Lab activities is the production of high-quality and unique biologicals that are not available from commercial sources. On average, the Unit performs around 50 processes last year, which include fermentation of bacteria, yeast, insect cells, and mammalian cells (in volumes ranging from 2 to 100 liters), and initial recovery and purification of biomolecules. The following are examples of processes performed: growth of bacteria such as Escherichia coli, (recombinant and native) various recombinant yeast strains such as Pichia pastoris. In addition, mammalian cells such as CHO, Hela, PG13 and HEK 293, insect cell such as Sf 9, for transient expression of recombinant proteins were propagated. The various products were needed for several collaborative research projects such as expression of P-glycoprotein (membrane protein transporter) from pichia pastoris (NCI), peptidoglycan from Bacillus anthraces (NIH clinical center) polysaccharides from E. coli (FDA) and production of human OLFM4 from HEK 293 cells. The lab continues its project together with the surgery branch of the NCI for improving the current method to produce gammaretroviral vectors needed for adoptive T cell therapy. The existing procedure is based on production from adherent packaging cells growing on stationary support which is a rate limiting step in this research. The modified process is based on growing the adherent cells on microcarriers in a stirred tank bioreactor. Using this methodology, we could produce infective particles in higher yield. The process was optimized and perfusion system which allow us to increase the cell number and the production yield was incorporated. Another approach that is currently at work is an alternative production based on lentiviral vector from cells growing in suspension.
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