Studying Molecular Interactions at the Single Bond Level with a Laminar Flow Chamber.

During the last decade, many investigators developed new methodologies allowing to study ligand-receptor interactions with unprecedented accuracy, up to the single bond level. Reported results include information on bond mechanical properties, association behaviour of surface-attached molecules, and dissection of energy landscapes and reaction pathways. The purpose of the present review is to discuss the potential and limitations of laminar flow chambers operated at low shear rates. This includes a brief review of basic principles, practical tips and problems associated with data interpretation. It is concluded that flow chambers are ideally suited to analyze weak interactions between a number of biomolecules, including the main families of adhesion receptors such as selectins, integrins, cadherins and members of the immunoglobulin superfamily. The sensitivity of the method is limited by the quality of surfaces and efficiency of the studied ligand-receptor couple rather than the hardware. Analyzing interactions with a resolution of a piconewton and a few milliseconds shows that ligand-receptor complexes may experience a number of intermediate binding states, making it necessary to examine the definition of association and dissociation rates. Finally, it is emphasized that association rates measured on surface-bound molecules are highly dependent on parameters unrelated to binding surfaces.