Nanopore-based kinetics analysis of individual antibody-channel and antibody-antigen interactions

Background: The UNO/RIC Nanopore Detector provides a new way to study the binding and conformational changes of individual antibodies. Many critical questions regarding antibody function are still unresolved, questions that can be approached in a new way with the nanopore detector. Results: We present evidence that different forms of channel blockade can be associated with the same antibody, we associate these different blockades with different orientations of "capture" of an antibody in the detector's nanometer-scale channel. We directly detect the presence of antibodies via reductions in channel current. Changes to blockade patterns upon addition of antigen suggest indirect detection of antibody/antigen binding. Similarly, DNA-hairpin anchored antibodies have been studied, where the DNA linkage is to the carboxy-terminus at the base of the antibody's Fc region, with significantly fewer types of (lengthy) capture blockades than was observed for free (un

• Type: Journal
• Year: 2007
• Where: BMCBI
• Authors: Stephen Winters-Hilt, Eric Morales, Iftekhar Amin, Alexander Stoyanov
• Keywords: Blockades, BMCBI 2007, Many Critical Questions, Nanopore Detector