Design And Preparation Of Enzyme Chip Applied For Pharmaceutical Screening

The impact of the new technologies is to accelerate the pace of drug discovery. Especially appearance of chip technology, which consumption of nanoliter volume makes low abundant samples able to be rapidly assayed. Protease is a kind of important drug target. In order to screen protease inhibitors on a great scale, a screening platform based on enzyme chip assay has been established and verified.Part â…  Design and preparation of chromogenic-based chip applied for screening elastase inhibitorsPulmonary emphysema is a main component of chronic obstructive pulmonary disease(COPD), currently the fourth leading cause of death in the United States and the only leading cause still increasing in prevalence. Neutrophil elastase is the crucial protease in the development of pulmonary emphysema. The finding stimulates interest in the search for agents with elastase inhibitory activity as a way of stopping, or at least slowing the development of emphysema. Here a chromogenic-based enzyme chip assay is described as a high-throughput format for screening elastase inhibitors.The detection principle of the assay is that elastase hydrolysises colorless polypeptide substrates and turns them into chromogenic products. Because the color of yellow of product isn't enough clear to limit the edge of a spot, bromochlorophenol blue(BPB) has been added into substrate solutions to emphasis profiles of spots. When enzymatic activity decrease, substrates can only be turned into blue in stead of yellow products. If enzyme retains its activity, yellow emitted by products will combine with blue of BPB, which makes the spots green. By measuring blue intensity of each spot, we can identify enzymatic activity of the corresponding site. A excellent linearity relationship has been observed between product concentrations and blue intensity of spots. The micro-system used for screening composes of five parts: glass slide, layer of agarose substratum, protease film, chemical arrays and substrate microarrays.