Immobilization strategy and characterization of hydrogel-based thin films for interrogation of ligand binding with staphylococcal enterotoxin B (SEB) in a protein microarray format

The preparation and characterization of 3-D hydrogel based thin films that employed a novel chemical immobilization strategy for protein attachment in a microarray format was discussed. Three-dimensional protein microarrays were printed using a noncontact microdispensing system that eliminated contact with the hydrogel substrate and minimized the potential for cross-contamination. Staphylococcal enterotoxin B (SEB) was used as the model protein and demonstrated antigenicity by a fluorescence-labeled anti-SEB antibody. The microfabricated 3-D hydrogel combined with chemical immobilization strategy provided reproducuble functional substrates for the immobilization of proteins with minimal background interference.