Kinetics of receptor and virus interaction and receptor-induced virus disruption: Methods for study with surface plasmon resonance

Abstract:

We have used BIAcore to analyze the kinetics of the interaction between rhinovirus and soluble intercellular adhesion molecule-1 (sICAM-1). Human rhinovirus serotype 3 (HRV3) was immobilized in the carboxymethylated dextran of the sensor chip, and sICAM-1 expressed with baculovirus was injected through the rhinovirus surface. sICAM-1 bound specifically to HRV3. The virus remained intact in the surface after 12 successive cycles of association and dissociation at 20°C. The association rate was slow (720 M−1 s−1) and the dissociation rate was moderate (1.8 × 10−3 s−1) for protein interaction. A dissociation constant (KD) of 2.5 (±0.18) μM was obtained from the kinetic constants. A slightly higher KD of 7.2 μM was obtained when equilibrium between virus and soluble receptor was reached in solution. At 30°C, binding of sICAM-1 disrupted HRV3, as monitored by a loss in resonance units.

Last updated on 10/06/2015