Patent ID: 11912984
Assignee: IONTAS LIMITED
Field: Pharmaceuticals (Chemistry)
Classification: CPC C  A | IPC A  C  G

Claim 0:
1. A method of producing a library of at least 105 mammalian cell clones containing DNA encoding a diverse repertoire of antibody molecules which differ in one or more complementarity-determining regions (CDRs), wherein the antibody molecules are displayed on surfaces of the mammalian cell clones, wherein the antibody molecules comprise or are linked to membrane anchors, and wherein the library contains donor DNA integrated at a fixed locus or at duplicate fixed loci, the method comprising
providing donor DNA molecules encoding antibody fragments comprising one or more CDRs, and mammalian cells with a genome size of greater than 2×107 base pairs,
introducing the donor DNA molecules into the mammalian cells and providing a site-specific nuclease within the mammalian cells, wherein the nuclease cleaves a recognition sequence in cellular DNA to create an integration site at which the donor DNA becomes integrated into the cellular DNA, integration occurring through DNA repair mechanisms endogenous to the mammalian cells, thereby creating recombinant cells containing donor DNA integrated at a fixed locus or at duplicate fixed loci in the cellular DNA, and
culturing the recombinant cells to produce clones,
thereby providing a library of at least 105 mammalian cell clones containing the donor DNA encoding the repertoire of antibody molecules, wherein the antibody molecules are displayed on the surface of their expressing cells.