Antigen binding specificity of antibodies patterned by microcontact printing

The ability to pattern functional biomolecules onto surfaces at the micrometer scale is critical to a large number of important biological methods. Since devices such as microarrays and biosensors, or surfaces for generation of complex cellular networks, require such "bioselective" surfaces, it is important to verify the functionality of the patterned biomolecules. Patterning of antibodies onto substrates by microcontact printing or soft lithography has been previously demonstrated, but the ability of these antibodies to selectively bind antigens has not been fully explored. This work re-examines the transfer of antibodies from elastomer stamps onto glass coverslips and documents the antigen-binding capabilities of these antibodies. Various microcontact print (MCP) applied antibodies are tested for selectivity in binding fluorescence-conjugated antigens or antigens detected with fluorescently labeled molecules, which are quantified by wide-field fluorescent microscopy, The fluorescence intensity of bound antigens is compared to that of MCP-applied controls, as well as to that of antibodies that were directly adsorbed to the glass substrates. The results indicate that MCP-applied antibodies, although they are less functional immunologically than are those that have been directly bath-adsorbed onto substrates, maintain their selective binding properties.