Patent ID: 11926818
Assignee: IONTAS LIMITED
Field: Pharmaceuticals (Chemistry)
Classification: CPC C  A | IPC A  C  G

Claim 1:
2. A method of screening for a binder that recognizes a target, wherein the binder comprises or is linked to a membrane anchor, the method comprising:
providing donor DNA molecules encoding a diverse repertoire of binders, and mammalian cells,
introducing the provided donor DNA molecules into the cells and providing a site-specific nuclease within the 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 cells, thereby creating recombinant cells containing donor DNA integrated at a fixed locus or at duplicative 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 binders,
culturing cells of the library to express the binders, wherein the binders are displayed on the surface of their expressing cells,
exposing the binders to the target, allowing recognition of the target by one or more cognate binders, if present, and
detecting target recognition by the one or more cognate binders using a target molecule that is recognized by the one or more cognate binders and that is either directly or indirectly labelled, or directly or indirectly immobilized on a solid surface, and
optionally further comprising recovering cells of a clone containing DNA encoding the cognate binder.