The Building Blocks of Discovery – Research Tools

Mouse models, cell lines, reagents, and antibodies—these are all research tools which are crucial to the progression of scientific research every day at Emory University, one of the nation’s leading universities that received over $550 million in extramural funds in 2016 alone. When important research tools are not available elsewhere, some labs at Emory University specialize in making the necessary tools to further their research goals and to help the research community as a whole.

This is the case for David Pallas, PhD, Associate Professor of Biochemistry, whose lab has specialized in making high quality antibodies. He started creating research tools 30 years ago for the purpose of progressing his research and many of the antibodies that his lab has generated are currently licensed to companies that provide research tools for the scientific community.

“Cells infected with this tumor virus become cancerous when the oncogenes are expressed,” Pallas clarifies as he explains his research on cancer causing oncogenes. His goal was to utilize these cancer causing viral oncogenes to study the affected signaling pathways in cancer. This is when he realized the limitations of existing research tools—namely the antibodies.

“Antibodies allow us to investigate a protein’s binding partners, what it interacts with, and where it goes in the cell,” Pallas explains. This would be useful for investigating each step in the signaling pathway and furthermore identify downstream targets that are modified by these cancer-causing oncogenes. However, at that time, the antibodies weren’t very specific and attached to a variety of targets.

To create the antibodies he needed, he began to develop methodology to produce highly specific monoclonal and polyclonal antibodies. He developed methods for purifying difficult to purify proteins as well as methods to hyperimmunize rabbits and mice to very conserved antigens, which can be difficult. “To produce monoclonal antibodies we then used methods pioneered by others, collecting the immune cells out of a mouse, for example, and fusing them to a cancer cell to make immortal antibody producing cells,” Pallas explains. These cells then produce identical monoclonal antibodies ensuring each antibody’s quality and specificity. These antibodies are also high titer, which means that only small volumes of antibody are needed for each Western Blot, immunofluorescence, or flow cytometry procedure.

The antibodies David Pallas created guided him through his investigations of the signaling cascade and its downstream targets. To this day, his lab has licensed 17 antibodies and a portion of the royalties earned by Emory come back to his lab to support research projects in the lab. These licensed research tools are furthermore able to benefit the scientific community as a whole. Other labs appreciate the quality of his polyclonal and monoclonal antibodies for their high specificity and ability to produce strong, clear results and each use of the antibody helps further scientific progress.

Among his most significant inventions are monoclonal antibodies for the PP2A (protein phosphatase 2A) catalytic and regulatory subunits. These antibodies could be used to quantify levels of PP2A in cells, for example. PP2A, a tumor suppressor implicated in cancer by the Pallas lab in 1990, is able to target and turn off certain oncogenes. This crucial role makes it a popular therapeutic target for cancer research.

As part of his research, Pallas studies the role of PP2A methylation. This is the process by which a highly specialized enzyme known as methyltransferase attaches a methyl group onto PP2A, enabling PP2A to bind to certain regulatory subunits that affect growth control and apoptosis and control the phosphorylation of a protein central to Alzheimer’s Disease pathology. Dr. Pallas’ lab has developed monoclonal antibodies capable of determining the level of methylation on PP2A. These reagents are thus significant not only to cancer research, but also relevant to Alzheimer’s and neural developmental research. As Pallas continues through his investigations of knocking out the methyltransferase gene in conditional knockout mice, the research community excitingly looks for the potential signaling pathways or therapeutic cancer targets that could be discovered.

To find a complete list of research tools available at Emory go to this web page.