Large-scale production and purification of biological compounds

Shiloach, Joseph

Abstract

A significant portion of the Labs activities is the production of high-quality and unique biologicals that are not available from commercial sources. On average, the Unit performs between 100 to 150 processes per year, which include fermentation of bacteria, yeast, insect cells, and mammalian cells (in volumes ranging from 2 to 200 liters), and recovery and purification of biomolecules, such as proteins and polysaccharides, from different sources. The following are examples of processes performed: growth of bacteria like Escherichia coli, (recombinant and native), Salmonella, Shighella, Staphylococcus, Haemophilus influenzae , Bronchaseptica sp, Micobacteria sp, and various recombinant yeast strains like Saccharomyces cerevisiae and Pichia pastoris. In addition, mammalian cells such as HeLa, CHO, and HEK 293, and insect cells like Sf 9, and High five for transient expression of recombinant proteins were propagated. Several proteins were produced and purified in gram quantities. Examples are: recombinant Clostridium difficle toxin A, recombinant human adiponectin, recombinant human alpha1 proteinase inhibitor and recombinant Diphtheria toxin. The various products were needed for a number of collaborative research projects: human adiponectin was made for biological studies, human alpha 1 proteinase inhibitor for glycosylation studies, the bacterial toxins for vaccine development and several proteins were produced for structural studies among them Toll proteins.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
• Type: Scientific Cores Intramural Research (ZIC)
• Project #: 1ZICDK015500-48
• Application #: 7970066
• Support Year: 48
• Fiscal Year: 2009
• Total Cost: $323,620

Institution

• Name: National Institute of Diabetes and Digestive and Kidney Diseases

Publications

Mulangu, Sabue; Alfonso, Vivian H; Hoff, Nicole A et al. (2018) Serologic Evidence of Ebolavirus Infection in a Population With No History of Outbreaks in the Democratic Republic of the Congo. J Infect Dis 217:529-537

Inwood, Sarah; Xu, Hui; Black, Mary A et al. (2018) Continuous production process of retroviral vector for adoptive T- cell therapy. Biochem Eng J 132:145-151

Azurmendi, Hugo F; Battistel, Marcos D; Zarb, Jasmin et al. (2017) The ?-reducing end in ?(2-8)-polysialic acid constitutes a unique structural motif. Glycobiology 27:900-911

Yoon, Dennis J; Chen, Kevin Y; Lopes, André M et al. (2017) Mathematical modeling of mutant transferrin-CRM107 molecular conjugates for cancer therapy. J Theor Biol 416:88-98

Qiang, Xiaoling; Liotta, Anthony S; Shiloach, Joseph et al. (2017) New melanocortin-like peptide of E. coli can suppress inflammation via the mammalian melanocortin-1 receptor (MC1R): possible endocrine-like function for microbes of the gut. NPJ Biofilms Microbiomes 3:31

Esser, Lothar; Zhou, Fei; Pluchino, Kristen M et al. (2017) Structures of the Multidrug Transporter P-glycoprotein Reveal Asymmetric ATP Binding and the Mechanism of Polyspecificity. J Biol Chem 292:446-461

Xu, Xinhui; Hicks, Caitlin; Li, Yan et al. (2014) Purified cell wall from the probiotic bacterium Lactobacillus gasseri activates systemic inflammation and, at higher doses, produces lethality in a rat model. Crit Care 18:R140

Negrete, Alejandro; Pai, Amrita; Shiloach, Joseph (2014) Use of hollow fiber tangential flow filtration for the recovery and concentration of HIV virus-like particles produced in insect cells. J Virol Methods 195:240-6

Wang, Hailong; Li, Yongjiang; Truong, Lan N et al. (2014) CtIP maintains stability at common fragile sites and inverted repeats by end resection-independent endonuclease activity. Mol Cell 54:1012-21

Xiao, Su; White, Jim F; Betenbaugh, Michael J et al. (2013) Transient and stable expression of the neurotensin receptor NTS1: a comparison of the baculovirus-insect cell and the T-REx-293 expression systems. PLoS One 8:e63679