Residence time prediction in magnetically controlled biomolecular local rebinding-dissociation kinetics

Abstract

The residence time of drug-target conjugates is a critical factor in drug screening and efficacy prediction. The local rebinding-dissociation kinetics gives insights into in-vivo drug-target interactions. A magnetic torque system (MTS) is designed to observe rebinding-dissociation kinetics for predicting residence time. The system utilizes an alternating magnetic field (AMF) to manipulate the magnetization motion of magnetically labeled biomolecules and the forces acting upon biomolecular bonds. The motion, sensed by a quartz crystal microbalance (QCM), reflects biomolecular interactions occurring at the particle surface. Meanwhile, the motion facilitates the separation of dissociated molecules from the surface, thereby obviating the necessity for fixed and mobile phases in common kinetics observations. The constant and static solution environment minimizes reagent consumption. The MTS was utilized to observe the local rebinding-dissociation of antibodies (PAB and MAB) to magnetic beads (MB) and to HER2 receptors. The residence times recorded by the MTS were larger than the results obtained via SPR method, due to the occurrences of rebinding-dissociation kinetics. Interaction behaviours can be meticulously regulated for varying affinities by modulating the intensity of magnetic field. A high intensity field (400 Oe) was applied for strong binding between antibody-MB (biotin-streptavidin), and a low intensity field (300 Oe) was applied for weak antigen-antibody interactions. An increase in AMF strength enhanced dissociation, with a shift from 300 Oe to 400 Oe resulting in a 1 ∼ 4-fold reduction in residence time. Overall, the MTS provides an interactive and customizable perspective on kinetics observations.