The examination of antibody structure via
single molecule Förster resonance energy transfer (FRET). This technique
involves incorporating two dye molecules into a protein of interest at
known locations. The distance between the two dye molecules can be
determined based on their relative fluorescence intensities. By
examining the fluorescence from individual proteins labeled with the dye
molecules, a histogram of the observed distances between the dyes can
be formulated. This histogram allows the presence of multiple protein
conformations to be detected. The primary goal of Southern's research is
to determine whether or not distinct antibody conformations
exist. Southern is interested in antibody structure because it can be
affected by mutations in the antibody, and can influence the ability of
an antibody to instigate an immune response. In some cases, increasing
an immune response is the desired effect; in others, such as autoimmune
disorders, diminishing an immune response is preferred. Studying the
structure of antibodies and how this structure is affected by mutations
or the removal of sugars bound to antibodies can shed light on the role
these structural changes play in generating an immune response.
Effect of Sugar Removal on the Structure of the Fc region of an IgG
Antibody as Observed with Single Molecule Förster Resonance Energy
M. T. Kelliher, R. D. Jacks, M. S. Piraino, C. A. Southern, Molecular Immunology, 2014, 60, 103. link
A Comparison of Förster Resonance Energy Transfer Analysis Approaches for Nanodrop Fluorometry,”
M. T. Kelliher, M. S. Piraino, M. E. Gemoules, C. A. Southern, Analytical Biochemistry, 2013, 441, 44. link
and Fluorescence Studies of Affinity Maturation in Related Antibodies,”
T. Pauyo, G. J. Hilinski, P. T. Chiu, D. E. Hansen, Y. J. Choi, D. I.
Ratner, N. Shah-Mahoney, C. A. Southern, P. B. O’Hara, Molecular Immunology, 2006, 43, 812. link
State and Multifrequency Phase Fluorometry Studies of Binding Site
Flexibility in Related Antibodies,” G. S. Mohan, P. T. Chiu, C. A.
Southern, P. B. O’Hara, Journal of Physical Chemistry A, 2004, 108, 7871. link
Consequences of Localized Electronic Excitation in Anthranilic Acid
Dimer,” C. A. Southern, D. H. Levy, J. A. Stearns, G. M. Florio, A.
Longarte, T. S. Zwier, Journal of Physical Chemistry A, 2004, 108, 4599. link
and Infrared Spectroscopy of Anthranilic Acid in a Supersonic Jet,” C.
A. Southern, D. H. Levy, G. M. Florio, A. Longarte, T. S. Zwier, Journal of Physical Chemistry A, 2003, 107, 4032. link