Interfacial immobilization of monoclonal antibody and detection of human prostate-specific antigen.

Antibody orientation and its antigen binding efficiency at interface are of particular interest in many immunoassays and biosensor applications. In this paper, spectroscopic ellipsometry (SE), neutron reflection (NR), and dual polarization interferometry (DPI) have been used to investigate interfacial assembly of the antibody [mouse monoclonal anti-human prostate-specific antigen (anti-hPSA)] at the silicon oxide/water interface and subsequent antigen binding. It was found that the mass density of antibody adsorbed at the interface increased with solution concentration and adsorption time while the antigen binding efficiency showed a steady decline with increasing antibody amount at the interface over the concentration range studied. The amount of antigen bound to the interfacial immobilized antibody reached a maximum when the surface-adsorbed amount of antibody was around 1.5 mg/m(2). This phenomenon is well interpreted by the interfacial structural packing or crowding. NR revealed that the Y-shaped antibody laid flat on the interface at low surface mass density with a thickness around 40 Å, equivalent to the short axial length of the antibody molecule. The loose packing of the antibody within this range resulted in better antigen binding efficiency, while the subsequent increase of surface-adsorbed amount led to the crowding or overlapping of antibody fragments, hence reducing the antigen binding due to the steric hindrance. In situ studies of antigen binding by both NR and DPI demonstrated that the antigen inserted into the antibody layer rather than forming an additional layer on the top. Stability assaying revealed that the antibody immobilized at the silica surface remained stable and active over the monitoring period of 4 months. These results are useful in forming a general understanding of antibody interfacial behavior and particularly relevant to the control of their activity and stability in biosensor development.

Multiple grasp-specific representations of tool dynamics mediate skillful manipulation.

Skillful tool use requires knowledge of the dynamic properties of tools in order to specify the mapping between applied force and tool motion. Importantly, this mapping depends on the orientation of the tool in the hand. Here we investigate the representation of dynamics during skillful manipulation of a tool that can be grasped at different orientations. We ask whether the motor system uses a single general representation of dynamics for all grasp contexts or whether it uses multiple grasp-specific representations. Using a novel robotic interface, subjects rotated a virtual tool whose orientation relative to the hand could be varied. Subjects could immediately anticipate the force direction for each orientation of the tool based on its visual geometry, and, with experience, they learned to parameterize the force magnitude. Surprisingly, this parameterization of force magnitude showed limited generalization when the orientation of the tool changed. Had subjects parameterized a single general representation, full generalization would be expected. Thus, our results suggest that object dynamics are captured by multiple representations, each of which encodes the mapping associated with a specific grasp context. We suggest that the concept of grasp-specific representations may provide a unifying framework for interpreting previous results related to dynamics learning.