Generation of binding molecules

Patent metadata

Field	Value
Publication number	US-2025230579-A1
Application number	US-202519170392-A
Country	US
Kind code	A1
Filing date	Apr 4, 2025
Priority date	Sep 26, 2011
Publication date	Jul 17, 2025
Grant date	—

Abstract

Provided are methods for efficiently and comprehensively screening antibody repertoires from B cells to obtain and produce molecules with binding characteristics and functional activities for use in human therapy.

First claim

1 . A method for producing a population of antibodies which bind to an antigen, said method comprising:
a) providing lymphoid tissue containing a population of B cells from a transgenic non-human animal immunized with said antigen; wherein said transgenic non-human animal expresses a single rearranged human antibody light chain variable region (VL) from a VL nucleic acid, wherein said transgenic non-human animal expresses a human antibody heavy chain variable region (VH) repertoire from a repertoire of VH nucleic acids, said B cell population expressing a repertoire of VHs and the single immunoglobulin light chain variable (VL) region repertoire, said B cell population thus expressing a limited immunoglobulin light chain variable (VL) region repertoire, b) obtaining the nucleic acids encoding antibody heavy chain variable (VH) regions from said lymphoid tissue, c) obtaining nucleotide sequences of the obtained nucleic acids of step b), d) determining the frequency of nucleotide sequences from step c) which encode VH regions, e) providing host cells, wherein each host cell comprises at least one vector comprising a nucleotide sequence encoding at least one VH region selected from those determined in step d) wherein each said host cell expresses the single rearranged human VL nucleic acid of step a) and said at least one VH region selected in step d), f) culturing said host cells from step e) and allowing for expression of said VL and VH regions, and g) obtaining said population of antibodies from the cultured host cells of step f) which bind said antigen.

2 . The method of claim 1 , wherein the method is without performing hybridoma generation between steps b) and c).

3 . The method of claim 1 , wherein the method is without use of display technology between steps b) and c).

4 . The method of claim 1 , further comprising subjecting a sample of said cultured host cells of step g) to at least one functional assay to confirm binding to said antigen, and selecting at least one cell that produces an antibody which binds said antigen.

5 . The method of claim 1 , wherein the transgenic non-human animal has been immunized with a nucleic acid encoding said antigen or with a protein form of said antigen.

6 . The method of claim 1 , wherein the transgenic non-human animal is a rodent.

7 . The method of claim 1 , wherein the transgenic non-human animal is a mouse.

Assignees

• Merus Nv

Inventors

• Throsby Mark
• Logtenberg Ton
• De Kruif John

Classifications

• C07K16/108

  Orthomyxoviridae (F), e.g. influenza virus · CPC title

• C07K2317/56

  variable (Fv) region, i.e. VH and/or VL · CPC title

• C07K2317/21

  from primates, e.g. man · CPC title

• C07K16/468 Primary

  Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies · CPC title

• C12Q1/6869

  Methods for sequencing · CPC title