A diverse collection of unique methodologies to synthesize and construct protein arrays for basic and clinical research in a high-throughput manner. These readily reproducible techniques can be applied to such applications as quantifying specific proteins of interest and discovering novel proteins, as well as to a variety of affinity substances, including antibodies, peptides, proteins, aptamers, and chemicals. The techniques can be used for biomarkers discovery, assay development, clinical sample testing, and building the next generation of molecular tools.