Large-scale production and purification of biological compounds

National Institute Of Diabetes And Digestive And Kidney Diseases

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Abstract

Summary: An important portion 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 100 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 Saccharomyces cerevisiae and Pichia pastoris. In addition, mammalian cells such as HeLa, CHO, MDCK and HEK 293, insect cell such Sf 9, and High five for transient expression of recombinant proteins were propagated. The various products were needed for a number of collaborative research projects: Expression of P-glycoprotein (membrane protein transporter) from pichia pastoris (NCI), expression of human NPP1 in HEK 293 cells (Yale university medical school), and peptidoglycan from Bacillus anthraces (NIH clinical center). The lab initiated a new project together with the surgery branch of the NCI for improvement of the method currently used for production of 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. Our aim is to improve the growth procedure by testing various methodologies using stirred tank reactor. Initial experiments were successfully conducted for the growth of the packaging cells in bioreactors