Preparation of libraries of protein variants expressed in eukaryotic cells and use for selecting binding molecules

a technology of protein variants and libraries, applied in the field of protein variant libraries expressed in eukaryotic cells and selection of binding molecules, can solve the problems of limited ability to screen the output, inability to resolve the extent of expression and the binding affinity of resultant clones, and limitations of the prokaryotic display system including phage display
US20200165597A1Inactive Publication Date: 2020-05-28IONTAS LTD
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Patent Information

Authority / Receiving Office
US · United States
Patent Type
Applications(United States)
Current Assignee / Owner
IONTAS LTD
Publication Date
2020-05-28
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to methods of producing eukaryotic cell libraries encoding a repertoire of binding molecules (“binders”), wherein the methods use a site-specific nuclease for targeted cleavage of cellular DNA to enhance site-specific integration of binder genes through endogenous cellular repair mechanisms. Populations of eukaryotic cells are produced in which a repertoire of genes encoding binders are integrated into a desired locus in cellular DNA (e.g., a genomic locus) allowing expression of the encoded binding molecule, thereby creating a population of cells expressing different binders.
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Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation of U.S. patent application Ser. No. 15 / 308,570, filed Nov. 2, 2016, which is a 35 U.S.C. § 371 application of International Application Serial No. PCT / GB2015 / 051287, filed May 1, 2015; which claims the benefit of Great Britain Application Number 1407852.1 that was filed on May 2, 2014. The entire content of the applications referenced above are hereby incorporated by reference herein.SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Oct. 31, 2016, is named 00130_003US1_SL.txt and is 92.9 kilobytes in size.FIELD OF THE INVENTION

[0003] This invention relates to methods of producing eukaryotic (e.g., mammalian) cell libraries for screening and / or selection of binding molecules such as antibodies. Libraries can be used to contain and display a divers...

Claims

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