Antibodies are Y-shaped, flexible proteins whose structures can be studied using Förster Resonance Energy Transfer (FRET) at the single-molecule level. Dye molecules must be attached to these proteins so as to carry out FRET studies of antibodies. In order to label the binding sites of an antibody, dye molecules were attached to a small molecule, or hapten, which the antibody binds to. Evidence for this binding was provided by ultraviolet-visible (UV-Vis) spectroscopy. To label the stem region of a humanized immunoglobulin G (IgG) antibody, the DNA for this antibody was mutated to introduce a cysteine residue to which dyes can be attached. In this research, the DNA was sequenced and checked to provide the desired sequence for protein production.
"Single-Molecule Fluorescence Studies of Glycosylated and Aglycosylated Antibodies,"
DePaul Discoveries: Vol. 1
, Article 11.
Available at: http://via.library.depaul.edu/depaul-disc/vol1/iss1/11