Antibody-antigen hybridization

There are several important advantages RPMAs have over antibody arrays and other proteomic techniques such as immunohis-tochemistry or tissue arrays. Antibody arrays usually require a second specific antibody, made in a different species, for each captured protein to be visualized in a manner analogous to enzyme-linked immunosorbent assays (ELISA). Therefore, it becomes difficult to simultaneously optimize the antibody-antigen hybridization conditions for so many antibodies at once present on antibody arrays while minimizing nonspecific cross-reactivity and ensuring that proteins over a wide range of concentrations can be quantitated in a linear fashion (14). Antibody arrays also consume or require much higher inputs of protein than reverse phase arrays. With antibody arrays. 

Built-in functional specific recognition and binding, e.g., antigen binding by antibodies or hybridization of nucleic acids ... 

Immunoaffinity chromatography (IAC), 6 400—402 12 137, 145 Immunoanalyzers, automated, 14 150 Immunoassay(s), 14 135-159. See also Immunoassay- DNA probe hybrid assays Immunoassay methods Immuno(bio)sensors antibody-antigen reaction, 14 136-138 basic technology in, 14 138-140 chemiluminescent, 14 150-151 classification of, 14 140-153 design of, 14 139-140 enzyme, 14 143-148 fluorescence, 14 148-150 highly specific, 14 153 historical perspective on, 14 136 microarrays and, 14 156—157 microfluidics in, 26 968—969 monoclonal versus polyclonal antibodies in, 14 152-153... 

Fig. 3. To produce monoclonal antibodies, antibody producing spleen cells troll] a mouse that has been immunized against an antigen am mixed with mouse myeloma cells. Under the proper conditions, pairs of the cells fuse to form antibody-producing hybrid-myeloma ( hybridoma ) cells, which can live indefinitely in culture. Individual hybridomas are grown in separate wells, and the antibodies they produce are tested against the antigen. When an effective cell line is identified, it is grown either in cuhure or in the body cavities of mice to produce large quantities of chemically identical, monoclonal antibodies (reprinted with permission from Olsen, 1986, p. 26, Copyright, National Academy Press)...

Due to the small volumes and feature sizes, reaction rates are found to be quite different in the microbiosensor system in comparison to their macro counter part. Most of this is due to the fact that diffusion is not the limiting factor in a reaction any longer. For example, the diffusion time of a particle with a diffusion coefficient = 10 m s is 15 min to travel a distance of 1 mm, but only 10 s to travel 100 /rm and only 0.1 s to travel 10/rm [37]. Therefore, DNA hybridization reactions, antibody-antigen binding events, and enzyme—substrate catalytic reactions take place in a fraction of the time required earlier. DNA hybridization can be accomplished in a matter of seconds in a microchannel system, while it takes in the order of an hour when employing standard Northern or Southern Blotting techniques with a piece of nylon membrane soaking in several milliliters of hybridization solution. 

Chimeric antibodies are hybrid molecules combining the antigen-specific variable domain of the mouse antibody fused to the constant regions of a human IgG molecule (Fig. 1.4). This reduces the risk of immunogenicity somewhat, and the human Fc domain prolongs the serum half-life and is more effective in triggering the effector systems of complement and Fc receptors. 

For ARBf i-type, when the surface effect is ignored, t = 1.5 from the Smoluchowski equation, and t = 1.4 0.15 from the antibody antigen reaction [17]. The general type of bonding probability [18, 19] (Table 2) is a hybrid of the RA -type and ARBf i-type, and can deal with die aggregation process that is between DLA and RLA. 

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