Towards an Alternative for Antibodies: Construction and Characterization of a Large Combinatorial Library of Diverse Binding Proteins

摘要

The goal of this to create a large and very diverse population of binding proteins from which individual members can be selected on the basis of target specificity and be used as a substitute for antibodies in biosensor and other applications. Toward this end, phage display technology was used to create a collection of seventeen distinct combinatorial libraries in which the binding surfaces of several small, stable parent proteins were randomized. The completed collection contains approximately 4x10(exp 9) different protein variants. Because of the types of parent molecules chosen and because of the way the libraries were designed, the variants are expected to have more physical stability than antibody molecules and to be able to function in more severe types of environments. In a test of the general binding properties of the collection, the library pool was taken through four rounds of panning against 31 randomly chosen test compounds (20 proteins, 4 peptides and 7 small molecules) . Apparent binding proteins were observed against 22/31 (71%) of the compounds tested (17 proteins, 2 peptides and 3 small organic molecules) indicating the library collection is a useful source of receptor proteins.