Histidine engineered light chain antibodies and genetically modified non-human animals for generating the same

摘要

A genetically modified non-human animal is provided, wherein the non-human animal expresses an antibody repertoire capable of pH dependent binding to antigens upon immunization. A genetically modified non-human animal is provided that expresses a single light chain variable domain derived from a single rearranged light chain variable region gene in the germline of the non-human animal, wherein the single rearranged light chain variable region gene comprises a substitution of at least one non-histidine encoding codon with a histidine encoding codon. Methods of making non-human animals that express antibodies comprising a histidine-containing universal light chain are provided.

主权项

1. A method of generating an antibody that exhibits pH-dependent binding to an antigen of interest comprising:
(a) co-expressing in vitro in a host cell:
(i) a first immunoglobulin heavy chain nucleotide sequence comprising a heavy chain variable region sequence encoding a heavy chain variable domain of a first antibody, wherein the first antibody binds to the antigen of interest with a desired affinity at a neutral pH, wherein the heavy chain variable region sequence is generated in a non-human animal comprising in its germline genome a nucleic acid sequence comprising a single rearranged human immunoglobulin light chain variable region sequence that comprises a human germline Vκ1-39 or Vκ3-20 gene segment rearranged with a human germline Jκ gene segment and that is expressed in the antibody repertoire of the non-human animal, and(ii) an immunoglobulin light chain nucleotide sequence that is derived from the single rearranged human immunoglobulin light chain variable region sequence and is modified to comprise a substitution of at least one non-histidine codon with a histidine codon designed to express a histidine at a position selected from 105, 106, 107, 108, 109, 111 and a combination thereof (according to IMGT numbering), and (b) selecting a second antibody expressed in the host cell that is encoded by the first immunoglobulin heavy chain nucleotide sequence and the immunoglobulin light chain nucleotide sequence, wherein the second antibody retains the desired affinity for the antigen of interest at neutral pH and displays reduced binding to the antigen of interest at an acidic pH.