Abstract
A novel protein microarray technology, called high-density nucleic acid programmable protein array (HD-NAPPA), enables the serological screening of thousands of proteins at one time. HD-NAPPA extends the capabilities of NAPPA, which producesprotein microarrays on a conventional glass microscope slide. By comparison, HD-NAPPA displays proteins in over 10,000 nanowells etched in a silicon slide. Proteins on HD-NAPPAare expressed in the individual isolated nanowells, via in vitro transcription and translation (IVTT), without any diffusion during incubation. Here we describe the method for antibody biomarker identification using HD-NAPPA, including four main steps: (1) HD-NAPPA array protein expression, (2) primary antibodies (serum/plasma) probing, (3) secondary antibody visualization, and (4) image scanningand data processing.
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