Abstract
Nanoporous poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) (HEMA-EDMA) is used as a 3D mesh for spotting lipid arrays. Its porous structure is an ideal matrix for lipid ink to infiltrate, resulting in higher fluorescent signal intensity as compared to similar arrays on strictly 2D substrates like glass. The embedded lipid arrays show high stability against washing steps, while still being accessible for protein and antibody binding. To characterize binding to polymer-embedded lipids we have applied Streptavidin as well as biologically important biotinylated androgen receptor binding onto 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(cap biotinyl) (Biotinyl Cap PE) and anti-DNP IgE recognition of 2,4-dinitrophenyl[1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[6-[(2,4-dinitrophenyl)amino]hexanoyl] (DNP)] antigen. This approach adds lipid arrays to the range of HEMA polymer applications and makes this solid substrate a very attractive platform for a variety of bio-applications.
Highlights
Starting with the creation of high density peptide microarrays in the 1990s [1] the development of arrays with DNA molecules and antibodies has produced a range of platforms for functional determination studies of molecules [2,3,4]
To determine if lipids deposited on nanoporous HEMAEDMA polymer could be used to generate functional arrays we selected lipid–protein pairs applied in previous settings: BiotinCap-PE and streptavidin labeled with Cy3 dye (STV-Cy3) as a simple protein model; and DNP-cap-PE with anti-DNP IgE as a model for allergen/antibody recognition
We present a novel platform for binding studies of functional molecules based on lipid arrays embedded in a nanoporous hydrophilic polymer substrate
Summary
Starting with the creation of high density peptide microarrays in the 1990s [1] the development of arrays with DNA molecules and antibodies has produced a range of platforms for functional determination studies of molecules [2,3,4]. To determine if lipids deposited on nanoporous HEMAEDMA polymer could be used to generate functional arrays we selected lipid–protein pairs applied in previous settings: BiotinCap-PE and streptavidin labeled with Cy3 dye (STV-Cy3) as a simple protein model; and DNP-cap-PE with anti-DNP IgE as a model for allergen/antibody recognition. For a more complex protein, the lipid/HEMA-EDMA substrate system was characterized for the binding properties of biotinylated androgen receptor (ARbiot) [18] onto Biotincap-PE arrays in the presence of a hormone.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
