Synthetic gene libraries

Details Synthetic gene libraries Synthetic gene libraries

C - Chemistry, Metallurgy

12

P

C12P 21/00 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2125240

A method for the simultaneous randomization of multiple and specific regions of a protein is described. This method also controls the degree to which this randomization occurs. Targeted sequences within a protein are partially randomized by the precise randomization of synthetic deoxyribooligonucleotides. Genes encoding proteins are assembled from the randomized oligonucleotides using a ligase reaction and are expressed on bacteriophage surfaces for selection of novel activities. The method can be used for randomizing antibody complementarity determining regions or framework regions for enhancing the affinity of the mutated antibodies. In one example, mutant single-chain antibodies, specific for the Salmonella serogroup B O-antigen, with 100-fold better binding, relative to the wild-type, were isolated using the invention.

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