Phage antibody fragments library combining a single human light chain variable region with immune mouse heavy chain variable regions

Abstract

We describe the construction of a phage antibody fragments library which combines, in a single cloning step, a synthetic human light chain variable region (V L) with a diverse set of heavy chain variable regions, from a mouse immunized with the prostate specific antigen (PSA). Despite V L restriction, selection from this library rendered two different single chain Fv antibody fragments, specifically recognizing PSA. The human V L, used as a general partner for mouse heavy chains, was constructed by linking the germline A27 gene and the J K 1 minigene segment, both of which are prominently involved in human antibody responses. Our approach offers a fast and simple way to produce half-human molecules, while keeping the advantage of immunizing animals for high affinity antibodies.