A Biomimetic Approach for the Creation of Two-Dimensional Microscale Surface Patterns: Creation of Isolated Immunological Synapses

Figure 1

Sequence of photopatterning and chemical treatment steps for binding antibodies and PEG to create surface APC mimics. The steps are thoroughly discussed in the text. Briefly, Step  1 shows the first layer of photoresist patterning followed by subsequent glutaraldehyde conjugation to exposed amine (NH₂) groups. Step  2 is the second photopatterning (following removal of the first layer) with subsequent thiol (SH) conjugation to exposed NH₂ groups. In Step  3, PEG is bound to aldehyde (CHO) groups and free SH groups are biotinylated. Step  4A demonstrates IgG conjugation to exposed NH₂ groups and biotinylated-IgA binding to surface-bound streptavidin-Ac. Antibody recognition of specific antigens enables fluorescently conjugated antibodies to specifically bind active IgG and IgA. Step  4B shows a similar binding of two fluorescently-labeled IgGs, which create an artificial IS in this geometry.