Joseph Shiloach, Ph.D.
Building 14A, Room 173
14 Service Road West
Bethesda, MD 20814
The purpose is to come up with efficient methodologies for the production of various biological compounds, such as proteins and viruses, from different types of biological sources, especially bacteria and mammalian cells. These biological products are being used currently for clinical and structural studies directed toward the development of effective new drugs and vaccines.
My laboratory’s main interest is the production and purification of biological products from both prokaryotes and eukaryotes of native and recombinant strains. Our work involves all aspects of this process including research, development, and production. The laboratory includes a pilot production facility that is well equipped with recovery and purification equipment and bioreactors of various sizes and configurations. These bioreactors are used for the propagation of biological systems such as mammalian cells, insect cells, bacteria, yeast, and fungi. The biological products we make support clinical and structural studies.
We conduct research and process development based on growth optimization, production scale-up, and product-recovery processes. We concentrate on protein production and purification, with an emphasis on scaling up. The aim of our research studies is to achieve better understanding of the growth behavior and metabolism of both E. coli and several mammalian cell lines. The objective is to overcome specific difficulties in the growth and production process by modifying the properties of the producers. This work involves gene transcription and expression analysis as well as cell transfection and mutation. Our recent achievements are as follows:
- We identified genes involved in mammalian cell adhesion, which allows us to modify the way the cells grow. The gene Siat7e was overexpressed in the adherent cells MDCK and, as a result, the cells were able to grow in suspension. These cells were then utilized for the preparation of influenza vaccine replacing the current influenza vaccine production, which is based on using fertilized eggs.
- We identified microRNAs that are involved in controlling apoptosis onset in mammalian cells, allowing us to extend the growth phase of the cells and potentially enhance the production of biological compounds. Inhibition of the microRNA 466h increased the expression levels of anti-apoptotic genes and increased the cell viability.
- We improved glucose utilization by E. coli through the over-expression of small RNA. This was responsible for glucose transport, allowing the bacteria to grow to high density. Over expression of the small RNA sgrS in E. coliK strain reduced the level of the glucose transporter PTSG and allowed the bacteria to increase the glucose utilization yield, reduce acetate production, and to increase the bacteria concentration and recombinant protein production.
Applying our Research
Improved methodologies for the production and purification of biological compounds intended for clinical research will provide researchers with the materials needed for the evaluation and testing of potential new drugs and vaccines against various diseases.
- Adjunct Professor, Johns Hopkins University, Department of Chemical and Biochemical Engineering, 2004
- Research Associate, Tufts University, New England Enzyme Center, 1975-1978
- Ph.D., Hebrew University, Israel, 1975
Baez A, Shiloach J. Increasing dissolved-oxygen disrupts iron homeostasis in production cultures of Escherichia coli. Antonie Van Leeuwenhoek. 2017;110(1):115-124.
Robbins JB, Schneerson R, Kubler-Kielb J, Keith JM, Trollfors B, Vinogradov E, Shiloach J. Toward a new vaccine for pertussis. Proc Natl Acad Sci U S A. 2014;111(9):3213-6.
Xiao S, Chen YC, Buehler E, Mandal S, Mandal A, Betenbaugh M, Park MH, Martin S, Shiloach J. Genome-scale RNA interference screen identifies antizyme 1 (OAZ1) as a target for improvement of recombinant protein production in mammalian cells. Biotechnol Bioeng. 2016;113(11):2403-15.
Chu C, Bottaro DP, Betenbaugh MJ, Shiloach J. Stable Ectopic Expression of ST6GALNAC5 Induces Autocrine MET Activation and Anchorage-Independence in MDCK Cells. PLoS One. 2016;11(2):e0148075.
Xiao S, Chen YC, Betenbaugh MJ, Martin SE, Shiloach J. MiRNA mimic screen for improved expression of functional neurotensin receptor from HEK293 cells. Biotechnol Bioeng. 2015;112(8):1632-43.
Related Scientific Focus Areas
Molecular Biology and Biochemistry
Genetics and Genomics
This page was last updated on December 3rd, 2015