MAY 04, 2016 01:00 PM PDT

Mammalian in vitro translation (cell-free) systems for rapid protein expression and production of therapeutic proteins

Speakers
  • Research Scientist, Thermo Fisher Scientific
    Biography
      Dr. Jensen is a Research Scientist at Thermo Fisher Scientific in the Cell Free Protein Expression group. Exciting research includes using our 1-Step CHO High Yield IVT system to express proteins and biologics on demand in collaboration with DARPA (Defense Advanced Research Projects Agency; for more information please see http://magazine.umbc.edu/umbc-magazine-winter-2016/making-medicine-mobile/ ). Previous work experiences include a Senior Scientist position at Sigma Aldrich developing tools for their shRNA library and two postdoctoral positions at Washington University in St. Louis studying Parkinson's disease and Diabetes.
    • Professor, Chemical & Biochemical Engineering, University of Maryland
      Biography
        Govind Rao is Professor of Chemical & Biochemical Engineering at the University of Maryland, Baltimore County. He obtained his Bachelor of Technology degree in Chemical Engineering from the Indian Institute of Technology, Madras in 1984. His Ph.D. degree was obtained from Drexel University in Chemical Engineering in 1987. He has been a faculty member at UMBC since 1987 and has served as Department Chair from 2000-2006. In 2006, he founded the Center for Advanced Sensor Technology (CAST) and has been serving as its Director since its inception.

        Dr. Rao's research is targeted towards disruptive innovation, where the goal is to create paradigm shifts in the state-of-the-art. He has focused on applications of fluorescence spectroscopy to create novel low-cost sensors for bioprocess, biomedical and environmental applications. His lab has developed next-generation sensors for low-cost non-invasive monitoring of oxygen, pH and pCO2 in bioreactors. In addition, novel sensors for glucose and glutamine have been developed. These sensors have led to a paradigm shift in bioprocess technology, as they collectively enable high throughput bioprocessing and are ideal for single use. Recently, the Rao lab has developed non-invasive sensor technology for neonatal monitoring. The current focus in the CAST is a major effort to develop next generation bio-manufacturing technology and is aimed at producing protein-based therapeutics at the point-of-care. A major effort at CAST is the application of sensor technology to reduce healthcare costs and close disparity gaps by making innovative low-cost devices for use in low-resource settings. As part of this strategy, technology transfer and partnerships have been set up with entities in low-resource settings. He has published over 160 papers in professional journals. His funding has come from several diverse sources including NSF, NIH, JDRF, DARPA, FDA, ONR and from several companies.

      Abstract:
      In vitro translation (IVT) using mammalian cell extracts is a quick and convenient alternative to in vivo mammalian protein expression. Thermo Fisher has developed IVT systems from two mammalian cell lines, HeLa and CHO, which enable rapid expression of genes from mammals, bacteria or protozoa. The Thermo Scientific 1-Step Human Coupled,  1-Step Human High-Yield, and  1-Step CHO High-Yield IVT kits allow rapid protein expression through the addition of a DNA of interest to a mixture of HeLa or CHO cell lysate and reaction mix followed by incubation for 1-6 hours at 30°C. For optimal expression in both IVT systems a series of vectors (pT7CFE1) with a variety of purification tags including Gateway™ compatible vectors for high throughput applications have been engineered. Thermo Fisher will present data demonstrating expression, purification, and activity of several different proteins using both the HeLa and CHO IVT kits. Results indicate that highly functional proteins can be produced in milligram quantities for a variety of downstream applications. Further, a new protein service option will be discussed encompassing gene synthesis, IVT expression, and purification with turnaround time less than 10 business days.

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