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Principal Investigator: Joseph Shiloach
Organization: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND KIDNEY DISEASES
Fiscal Year: 2021
Award: $531,271
Funding agency: National Institute of Diabetes and Digestive and Kidney Diseases
Summary:
An essential part of the Lab activities is production of unique biologicals that are not available from commercial sources at a suitable quality, for basic and clinical research. During the last year the Lab performed several processes, which include propagation of bacteria, and mammalian cells (in shake flasks and different size bioreactors) and executed initial recovery and purification of biomolecules. The following are examples of processes performed: growth of bacteria such as Escherichia coli and nonpathogenic Bacillus anthracis, yeast such as Pichia pastoris mammalian cells such as CHO, PG13 and HEK 293, for expression of recombinant proteins. The various products were needed for different collaborative research projects such as expression of P-glycoprotein (membrane protein transporter) from Pichia pastoris (NCI), peptidoglycan from Bacillus anthracis (NIH clinical center) and polysaccharides from E. coli (FDA). The Lab was heavily involved, together with Professor Andrew Pekosz, Department of Molecular Microbiology & Immunology Johns Hopkins University Bloomberg School of Public Health, in the production and initial purification of mg quantities of different Covid 19 proteins; especially different spike proteins and different receptor binding domain (RBD) which were needed for clinical evaluations. The production was done by expressing the covid proteins from HEK 293 cells growing in defined media. The process was associated with scaling up the protein expression with an emphasis on downstream recovery by using tangential flow filtration process. Another protein that the lab produced is Pigment epithelium-derived factor (PEDF) which was expressed from HEK Ebna cells and was needed for structural studies by the National Eye Institute. We also conducted earlier development of antibodies production process for Ebola research with the Vaccine Research center. Different types of media and transfection agents in different bioreactor configuration were tested.
Terms: <ABC20><ABCB1><ABCB1 gene><Ab response><Antibody Formation><Antibody Production><B anthracis><B. anthracis><Bacillus anthracis><Bacteria><Basic Research><Basic Science><Biocompatible Materials><Biological><Biomaterials><Bioreactors><Biotech><Biotechnology><Body Tissues><COVID><COVID-19><COVID19><CV-19><CV19><Carrier Proteins><Cell Body><Cell Density><Cells><Clinical Evaluation><Clinical Research><Clinical Study><Clinical Testing><CoV disease><Development><E coli><E. coli><EPC-1 protein><EPH-Like Tyrosine Kinase 1><EPHA3><EPHA3 gene><ETK1><Ebola><EphA3 Receptor><Ephrin Receptor EphA3><Escherichia coli><Filtration><Filtration Fractionation><GP170><Generalized Growth><Glycans><Growth><HEK Gene><HEK protein><HEK4><Hansenula><Human Embryo Kinase><Immunology><Insecta><Insects><Insects Invertebrates><Laboratories><Laboratory Research><MDR-1><MDR1><MDR1 Protein><Maintenance><Mammalian Cell><Membrane Protein Gene><Membrane Proteins><Membrane-Associated Proteins><Methodology><Methods><Microbiology><Molecular><Multidrug Resistance 1><Multidrug Resistance Gene-1><Multidrug Resistance Gene-1s><Multidrug Resistance Proteins><Multidrug Resistant Proteins><Murein><NIDDK><NIH><National Eye Institute><National Institute of Diabetes and Digestive and Kidney Diseases><National Institutes of Health><P-GP><P-Glycoprotein><P-Glycoprotein 1 Gene><PEDF><PGY-1 Protein><PGY1><Peptidoglycan><Pichia><Plants><Polysaccharides><Process><Production><Proteins><Public Health Schools><R-Series Research Projects><R01 Mechanism><R01 Program><Receptor Tyrosine Kinase HEK><Recombinant Proteins><Recombinants><Recovery><Research><Research Grants><Research Project Grants><Research Projects><Source><Structure><Surface Proteins><TYRO4><Testing><Tissue Growth><Tissues><Transfection><Transport Protein Gene><Transport Proteins><Transporter Protein><United States National Institutes of Health><Universities><Vaccine Research><Yeasts><anthracis><antibody biosynthesis><biological material><clinical center><clinical test><corona virus disease><corona virus disease 2019><coronavirus disease><coronavirus disease 2019><develop a vaccine><development of a vaccine><developmental><drug development><early PDL protein><early population doubling level cDNA-1 protein><early population doubling level protein><flasks><immunoglobulin biosynthesis><instrumentation><large scale production><microorganism><ontogeny><operation><pigment epithelium-derived factor><professor><protein expression><receptor binding><receptor bound><research clinical testing><scale up><structural biology><vaccine development><vaccine formulation>