Multi-component micro-aligned patterns have been constructed and 3-D spatial control for hydrogels fabricated by e-beam lithography has been demonstrated.  Cross-sectional analysis has confirmed that these microstructures have been produced such that an alkyne terminated 8-arm polyethylene glycol (Alkyne-PEG) is patterned inside a contiguous outer shell with 8-arm PEG of aminooxy functionality (AO-PEG) completing encapsulation of the hydrogel (Figure 1).  This strategy has been extended to achieve 3-D spatial control of immobilized proteins including two different enzymes, horse radish peroxidase (HRP) and glucose oxidase (GOx).  Activity of both immobilized enzymes has been confirmed in this encapsulated configuration via the reaction series between GOx and HRP.Figure 1: a) Top down view of Alkyne-PEG stained with azide functionalized AlexaFluor� 488 (white) patterned inside AO-PEG stained with maleimide functionalized coumarin (gray). b) xz through y transparency projection illustrating complete encapsulation of the alkyne-PEG core.  Scale bar is 20�m.

Multicomponent 3D protein arrays by electron beam lithography

243rd National Meeting (2012)

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