We can adapt and get different types of quantitative data, all analyzed on the same [Odyssey CLx] platform.
Dr. Michael Lee
University of Massachusetts Medical School
From a systems pharmacology perspective, Dr. Michael Lee is seeing cancer therapeutics in a whole new ‘infrared’ light. The Assistant Professor at the University of Massachusetts Medical School is analyzing biochemical responses of cancer cells to drugs, and attempting to emulate them computationally.
Growth factors and signaling networks involved in DNA damage are the focal points of research for Lee’s group, since these are largely thought to count for the therapeutic efficacies of common drugs. "I think what is really not understood is how processes like DNA damage interface with other signaling mechanisms to collectively control cell death. By looking at the cross-talk between the DNA damage response and EGFR signaling, we are trying to identify common nodes to see if we can predict how to use agents that target these pathways more effectively," says Lee. In one such study, the group developed a chemotherapy delivery system involving rewiring of signaling pathways1,2.
Quantitative Dynamic Range for Protein Expression Analysis
The Odyssey CLx Imager allows us to move forward in a more quantitative and a higher throughput manner...
Lee’s team creates temporal profiles of cellular protein levels or protein modifications in response to various drug concentrations and generates computational models of these large data sets. The lab is equipped with an Odyssey® CLx Imaging System that is routinely used for quantifying protein levels by Western blotting and Reverse Phase Protein Arrays.
Lee believes the real benefit of using the imager is its quantitative dynamic range. "You could make out a low band on film with high-quality enhanced chemiluminescence (ECL) that may not be visible using the Odyssey CLx System, but those signals are not really in the linear range of those reagents. During my Post-Doc when models [such as those that we work with] were being developed, they were collecting all of the data on film and the models were incredibly unreliable," he says. "For a lab that really cares about quantification, the data that would be produced by ECL on film is nowhere near quantitative enough or linear enough for our studies."
The instrument also found utility in various applications that their lab uses, including micro- and medium-throughput Western blotting, and In-Cell Western™ Assays. "We can adapt and get different types of quantitative data, all analyzed on the same [Odyssey CLx] platform," he says.
Increased Throughput With Direct Detection
In addition to the quantitative dynamic range, Lee feels the imager also provides better throughput. With film, stripping and reprobing is required for measuring a second or a third target. "It is just a waste of time, considering you can measure multiple targets at once on the Odyssey CLx System and it is probably more accurate to do this without stripping."
Members of the Lee lab love using the system. "The Odyssey CLx Imager allows us to move forward in a more quantitative and a higher throughput manner without completely ditching some of the things that are really nice about the old technology, like Western blotting which lets you physically see the data, at least the band sizes," says Lee.
Systems pharmacologists like Lee can efficiently profile protein levels for computational models using the Odyssey Infrared technology.
- Stephen W. Morton, Michael J. Lee, Zhou J. Deng, Erik C. Dreaden, Elise Siouve, Kevin E. Shopsowitz, Nisarg J. Shah, Michael B. Yaffe, and Paula T. Hammond (2014). A Nanoparticle-Based Combination Chemotherapy Delivery System for Enhanced Tumor Killing by Dynamic Rewiring of Signaling Pathways. Sci. Signal., 13 May 2014. Vol. 7, Issue 325, p. ra44. DOI: 10.1126/scisignal.2005261
- Michael J. Lee, Albert S. Ye, A lexandra K. Gardino, Anne Margriet Heijink, Peter K. Sorger, Gavin MacBeath, and Michael B. Yaffe (2012). Sequential Application of Anticancer Drugs Enhances Cell Death by Rewiring Apoptotic Signaling Networks. Cell, 11 May 2012, Vol 149, pg. 780-794.
- Dr. Geir Bjørkøy
University College of Sør-Trøndelag and CEMIR Centre of Excellence, NTNU
- Dr. Wendy Bollag
Georgia Regents University, Charlie Norwood VA Medical Center
- Dr. Jeremy Chambers
Florida International University, Miami, FL
- Dr. Paul Digard
University of Cambridge, UK
- Dr. Jane Gray
Cancer Research Institute, Cambridge, UK
- Manuela Gustafsson
Karolinska Institute's Department of Laboratory Medicine
- Dr. Oliver Hantschel
- Dr. Stefan Hüttelmaier
Martin Luther University
- Dr. Jelena Janjic
- Dr. Andrea Kasinski
- Dr. Sue Katz
Rogers State University
- Dr. Lisa Keyes
University of Florida College of Medicine
- Dr. Stanislav Kholmanskikh
Weill Cornell Medical College
- Dr. Patrick Kiely
University College Cork, Ireland
- Dr. Andreas von Knethen
Johann Wolfgang Goethe-University
- Dr. Michael Lee
University of Massachusetts Medical School
- Dr. Robert Lewis
University of Nebraska
- Dr. H. Charles Manning
- Dr. Debra Martin
Saint Mary's University of Minnesota
- Dr. Kevin Morano
The University of Texas Graduate School of Biomedical Sciences at Houston
- Dr. Rick Page
Miami University in Oxford, OH
- Dr. Nicolai Peschel
Universität Würzburg Department of Neurobiology & Genetics
- Dr. Wendy Picking
Oklahoma State University
- Dr. Martin Pomper
Johns Hopkins Medical School
- Dr. Vikash Reebye
Imperial College London, UK
- Dr. Rosemary Schuh
University of Maryland School of Medicine