Disclosed are a polyacrylamide-based method of fabricating surface-bound
peptide and protein arrays, the arrays themselves, and a method of using
the arrays to detect proteins and to measure their concentration, binding
affinity, and kinetics. Peptides, proteins, fusion proteins, protein
complexes, and the like, are labeled with an acrylic moiety and attached
to acrylic-functionalized glass surfaces through a copolymerization with
acrylic monomer. The specific attachment of GST-green fluorescent protein
(GFP) fusion protein was more than 7-fold greater than the nonspecific
attachment of non-acrylic labeled GST-GFP. Surface-attached GST-GFP (0.32
ng /mm.sup.2) was detectable by direct measurement of GFP fluorescence
and this lower detection limit was reduced to 0.080 ng/mm.sup.2 using
indirect antibody-based detection. The polyacrylamide-based surface
attachment strategy was also used to measure the kinetics of substrate
phosphorylation by the kinase c-Src. The surface attachment strategy is
applicable to the proteomics field and addresses denaturation and
dehydration problems associated with protein microarray development.