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Details of Grant 

EPSRC Reference: GR/S71491/01
Title: Basic Technology: Atomic force microscopy for real-time, qualitative monitoring of biological ligands in vivo
Principal Investigator: Lazarus, Dr CM
Other Investigators:
Miles, Professor MJ Napier, Professor RM
Researcher Co-Investigators:
Project Partners:
Department: Biological Sciences
Organisation: University of Bristol
Scheme: Standard Research (Pre-FEC)
Starts: 22 March 2004 Ends: 21 September 2005 Value (£): 77,234
EPSRC Research Topic Classifications:
EPSRC Industrial Sector Classifications:
Pharmaceuticals and Biotechnology
Related Grants:
Panel History:  
Summary on Grant Application Form
In addition to imaging dry surfaces at atomic resolution the atomic force microscope (AFM) can be used to assay biological macromolecules in solution. This application as a biosensor is based on the change in probe resonance frequency resulting from the increase in mass associated with the specific binding of a large ligand to its receptor immobilised on the tip. This research aims to vastly extend the utility of the AFM as a biosensor in biology and medicine with the capability to assay any small ligand (hormone, drug, metabolite) within individual (possibly living) cells and subcellular compartments. The concept to be proven involves loading a large molecule (protein) onto the probe tip via its specific interaction with immobilised ligand; on placing the tip in the test solution (e.g. a cell) containing free ligand the protein will be displaced from the tip, with concomitant loss of mass and change in resonance, at a rate dependent on the concentration of the free ligand. The system will be developed using the plant hormone abscisic acid as the small molecule to be assayed, with a single-chain antibody fragment (scFv) as the heavy protein to be displaced. Parameters to be optimised include the kinetics of interaction between the antibody and ligand, so a panel of scFvs with different defined kinetic constants will be isolated for evaluation by AFM.
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Organisation Website: http://www.bris.ac.uk