Provided are protein microarrays, their manufacture, use, and application.
Protein microarrays in accordance with the present invention are useful
in a variety preoteomic analyses. Various protein arrays in accordance
with the present invention may immobilize large arrays of proteins that
may be useful for studying protein-protein interactions to improve
understanding of disease processes, facilitating drug discovery, or for
identifying potential antigens for vaccine development. The protein array
elements of the invention are native or modified proteins (e.g.,
antibodies or fusion proteins). The protein array elements may be
attached directly to a organic functionalized mirrored substrate by a
binding reaction between functional groups on the substrate (e.g., amine)
and protein (e.g., activated carboxylic acid). Techniques for chemical
blocking of the arrays are also provided. The invention contemplates
spotting of array elements onto solid planar substrates, labeling of
complex protein mixtures, and the analysis of protein binding to the
array. The invention also enables the enrichment or purification, and
subsequent sequencing or structural analysis of proteins that are
identified as differential by the array screen. Kits including
protein-binding microarrays for proteomic analysis in accordance with the
present invention are also provided.