A wide range of biologically mediated methods and important applications of site-specific protein immobilization are discussed, through the culmination of a major multidisciplinary research effort spanning molecular biology. The study focused the attention on oriented protein immobilization methodology that allow defined attachment of proteins with uniform orientation at the bioactive site. The study include enzyme-mediated attachment to a small-molecule immobilizing ligand and the development of genetically encoded protein tags for immobilization. The advent of high-density array printing technology and improved methods for high-throughput production and purification of large number of proteins that allowed the preparation of protein microarrays. The development of proteome chip technology is used to probe an entire collection of proteins for specific function of biochemistry.