Partial plasma polymerization for coexistence of hydrophobic/hydrophilic area in several ten micrometer size is the typical technique for protein patterning. A hydrophobic hexamethyldisiloxane plasma-polymerized film (HMDS PPF) was deposited on a glass substrate and this surface was partially modified by subsequent nitrogen plasma treatment (hydrophilic surface, HMDS-N PPF) with a patterned shadow mask. An antibody protein (F(ab')2 fragment of anti-human immunoglobulin G) was selectively adsorbed onto the HMDS-N area and was not adsorbed onto the HMDS area. Distinct 8080 µm2 square spots surrounded by a non-protein adsorbed 80 µm-wide grid were observed. Then, when the protein modified by fullerene was used, the reversible patterning was obtained. This indicated that the modification by fullerene changed the hydrophilic nature of F(ab')2 protein to hydrophobic one, as a result, the modified protein was selectively adsorbed onto hydrophobic area.

Techniques for patterned modification of substrate surfaces are important for forming microarrays on protein chips. A hexamethyldisiloxane plasma-polymerized film (HMDS PPF) was deposited on a glass substrate and the resulting surface was partially modified by subsequent nitrogen plasma treatment with a patterned shadow mask. When surface adsorption of an antibody protein (F(ab')2 fragment of anti-human immunoglobulin G) was visualized by fluorescence microscopy, distinct 8080 µm2 square spots were observed, surrounded by a non-fluorescent 80 µm-wide grid. This pattern could be attributed to proteins selectively adsorbed onto the nitrogen plasma-treated regions but not onto the surface of pristine HMDS PPF. This provided a simple fabrication method of protein patterning.

Adsorption of antibody protein (anti-human IgG) onto plasma-polymerized thin films (PPF) with nanoscale thickness was characterized by atomic force microscopy (AFM) and quartz crystal microbalance (QCM). The PPF surface is very flat (less than 1 nm roughness) and its properties (charge and wettability) can be easily changed while retaining the backbone structure. We focus on two types of surfaces: one is the pristine surface of hexamethyldisiloxane (HMDS) PPF (hydrophobic) and the other is an HMDS PPF surface with nitrogen-plasma treatment (hydrophilic and positive-charged surface). The AFM image showed that the antibody molecules were densely adsorbed onto both surfaces and individual antibody molecules could be observed. The QCM profiles show a corresponding tendency with the AFM images. These results indicate that the plasma polymerized film can be the suitable biointerface for the application of biosensor and bioassay.

A high Tc superconducting quantum interference device (SQUID) magnetometer system is developed for the application to biological immunoassay. In this application, magnetic nanoparticles are used as magnetic markers to perform immunoassay, i.e., to detect binding reaction between an antigen and its antibody. The antibody is labeled with γ-Fe2O3 nanoparticles, and the binding reaction can be magnetically detected by measuring the magnetic field from the nanoparticles. Design and set up of the system is described, and the sensitivity of the system is studied in terms of detectable number of the magnetic markers. At present, we can detect 4106 markers when the diameter of the marker is 50 nm. Total weight of the magnetic nanoparticles becomes 520 pg in this case. An experiment is also conducted to measure antigen-antibody reaction with the present system. It is shown that the sensitivity of the present system is 10 times better than that of the conventional method using an optical marker. A one order of magnitude improvement of sensitivity will be realized by the sophistication of the present system.

We described short time span idiotype immune network reactions by rigorous mathematical equations. For each idiotype, we described the temporal changes in concentration of (1) single bound antibody, one of its two Fab arms has bound to the complemental receptor site on the B cell. (2) double bound antibody, both of its two Fab arms have bound to the complemental receptor sites on the B cell and (3) an immune complex which is a product of reaction among the antibodies. Stimulation and secretion processes of an antibody in the idiotype network were described by non linear differential equations characterized by the magnitude of cross-linking of the complemental antibody and B cell receptor. The affinity between the mutually complemental antibody and receptor was described by an weighted affinity matrix. The activating process was expressed by an exponential function with threshold. The rate constant for the linkage of the second Fab arm of an antibody was induced from the molecular diffusion process that was modified by the Coulomb repulsive force. By using reported experimental data, we integrated 60 non linear differential equations for the idiotype immune network to obtain the temporal behavior of concentrations of the species in hour span. The concentrations of the idiotype antibody and immune complex changed synchronously. The influence of a change in one rate constant extended to all the members of the idiotype network. The concentrations of the single bound antibody, double bound antibody and immune complex oscillated as functions of the concentration of the free antibody particularly at its low concentration. By comparing to the reported experimental data, the present computational approach seems to realize biological immune network reactions.