The Biotechnology Unit is responsible for the production and purification of biological material. The Unit undertakes a wide array of biological production tasks including large-scale growth of prokaryotic cells and eucaryotic cells, production of various compounds from microorganisms and eucaryotic dells, and purification of biological compounds especially proteins. These products, not available from commercial sources, are needed for clinical and structural studies. The Biotechnology Unit handles various production problems associated with growth and expression of recombinant and native products using both physiological and technical approaches. Efficient process development and production is possible since the Unit possesses two integrated elements: a fully operational multi-purpose pilot production facility and the capability of conducting research and process development. During the last year the unit performed approximately 200 different, large scale preparations including: 1, growing various microorganisms such as recombinant Escherichia coli, recombinant Saccharomyces cerevisiae, recombinant Pichia pastoris, mutant strains of Bortedella . pertussis, mutant strains of Corynebacteria diphtheria and several types of Salmonella and Shigella. 2, propagating large volumes of various mammalian cells; and 3, purifying gram quantities of diphtheria toxin CRM (Cross Reacting Material) and recombinant Pseudomonas aeruginosa exotoxin A. In the research and development arena the unit continues: 1, its effort on the development of more efficient methods for large scale protein recovery, performing experiments implementing expanded bed adsorption techniques for capturing proteins on an ion exchangers and chelating resins; 2, conducting research aimed at understanding the glucose metabolism of various recombinant E. coli strains in an effort to find out why certain strains are more suitable for foreign protein production; 3, exploring various method for propagation of anchorage dependent mammalian cells such as cellcubes , fluidized bed, and packed bed; and 4, understanding and improving of the process of recombinant protein expression from various yeast strains such as Pichia pastoris and Saccharomyces cerevisiae.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Intramural Research (Z01)
Project #
1Z01DK015500-37
Application #
6104999
Study Section
Special Emphasis Panel (LCDB)
Project Start
Project End
Budget Start
Budget End
Support Year
37
Fiscal Year
1998
Total Cost
Indirect Cost
City
State
Country
United States
Zip Code
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Phue, Je-Nie; Kedem, Benjamin; Jaluria, Pratik et al. (2007) Evaluating microarrays using a semiparametric approach: application to the central carbon metabolism of Escherichia coli BL21 and JM109. Genomics 89:300-5
White, Jim F; Grodnitzky, Justin; Louis, John M et al. (2007) Dimerization of the class A G protein-coupled neurotensin receptor NTS1 alters G protein interaction. Proc Natl Acad Sci U S A 104:12199-204
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Karnaukhova, Elena; Ophir, Yakir; Trinh, Loc et al. (2007) Expression of human alpha1-proteinase inhibitor in Aspergillus niger. Microb Cell Fact 6:34
Jaluria, Pratik; Betenbaugh, Michael; Konstantopoulos, Konstantinos et al. (2007) Application of microarrays to identify and characterize genes involved in attachment dependence in HeLa cells. Metab Eng 9:241-51
Kim, Chul-Hee; Pennisi, Patricia; Zhao, Hong et al. (2006) MKR mice are resistant to the metabolic actions of both insulin and adiponectin: discordance between insulin resistance and adiponectin responsiveness. Am J Physiol Endocrinol Metab 291:E298-305
Bell, Jessica K; Botos, Istvan; Hall, Pamela R et al. (2006) The molecular structure of the TLR3 extracellular domain. J Endotoxin Res 12:375-8
Trinh, Loc; Phue, Je-Nie; Jaluria, Pratik et al. (2006) Screen-less expanded bed column: new approach for the recovery and purification of a malaria transmission blocking vaccine candidate from Pichia pastoris. Biotechnol Lett 28:951-8
Shiloach, Joseph; Fass, Rephael (2005) Growing E. coli to high cell density--a historical perspective on method development. Biotechnol Adv 23:345-57

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