Richard Burgess, PhD
Position title: Oncology
Immunology Focus area: Preparation and use of mAbs in biochemical research, gentle immunoaffinity chromatography, immunochemistry, large-scale mAb purification, use of mAb affinity magnetic beads to isolate weakly bound protein complexes to identify new drug discovery targets.
Descriptive Title of Research: Just recently was awarded a 2-year SBIR grant with Salus Discovery LLC; Dave Beebe and Scott Berry in Salus and the Dept of Biomedical Engineering; and Shigeki Miyamoto and Richard Burgess labs in the Dept of Oncology. This grant is entitled “Seeing the unseen: Enhancing purification of labile protein complexes and cells with low surface marker expression”.
Research Description: In the late 80’s, we developed a new type of mAb that is ideal for the gentle immunoaffinity purification of labile proteins and protein complexes. These mAbs bind tightly under normal buffer conditions but release bound antigen when eluted with a very gentle buffer containing a salt, like ammonium sulfate, and a polyol, like propylene glycol. We call these mAbs polyol-responsive mAbs. More recently we have collaborated with Dave Beebe’s lab, using microfluidic methods, and can now rapidly purify proteins along with other proteins in a complex, even when the other proteins are only very weakly bound. We have concentrated on mAb-based immunoaffinity beads. The mAbs are bound to Protein G magnetic beads and mixed with crude extracts or samples to allow antigen binding. The beads are then drawn from the crude sample through a wash well to a final output well in seconds. This provides excellent washing but still retains weakly bound proteins, even those with binding half-lives of only a few seconds. In many ways this is like a super efficient sensitive Co-IP.
Link to Publications: (representative relevant references)
1) Thompson NE, Foley KM, Stalder ES, Burgess RR. Identification, production, and use of polyol-responsive monoclonal antibodies for immunoaffinity chromatography. Methods Enzymol 463, 475–494, 2009.
2) Stalder ES, Nagy LH, Batalla P, Arthur TM, Thompson NE, Burgess RR. The epitope for the polyol-responsive monoclonal antibody 8RB13 is in the flap-domain of the beta-subunit of bacterial RNA polymerase and can be used as an epitope tag for immunoaffinity chromatography. Protein Expr Purif 77(1), 26–33, 2011.
3) 2009. Berry SM, Chin EN, Jackson SS, Strotman LN, Goel M, Thompson NE, Alexander CM, Miyamoto S, Burgess RR, Beebe DJ. Weak protein–protein interactions revealed by immiscible filtration assisted by surface tension. Anal. Biochem. 2014 Feb;447:133–140.
Graduate Program Affiliations: In the past I have been a trainer in a number of programs, including Cancer Biology, Bacteriology, Cellular and Molecular Biology, Genetics. I am now a semi-retired professor emeritus of Oncology and no longer accept graduate students, but have a senior scientist, Nancy Thompson, who is an absolute expert on preparation and use of mAbs.
Departmental website: https://mcardle.wisc.edu/who-we-are/faculty/richard-r-burgess-phd Group website: http://mcardle.oncology.wisc.edu/burgess