Animal models and therapeutic molecules

Patent metadata

Field	Value
Publication number	US-9434782-B2
Application number	US-201314137902-A
Country	US
Kind code	B2
Filing date	Dec 20, 2013
Priority date	Jul 8, 2009
Publication date	Sep 6, 2016
Grant date	Sep 6, 2016

Abstract

The invention discloses methods for the generation of chimaeric human-non-human antibodies and chimaeric antibody chains, antibodies and antibody chains so produced, and derivatives thereof including fully humanized antibodies; compositions comprising said antibodies, antibody chains and derivatives, as well as cells, non-human mammals and vectors, suitable for use in said methods.

First claim

The invention claimed is:

1. A method for producing an antibody specific for a selected antigen, the method comprising
(a) providing a transgenic mouse whose genome comprises a homozygous chimeric immunoglobulin heavy chain (IgH) locus comprising unrearranged human IgH variable region gene segments at an endogenous mouse heavy chain IgH locus upstream of an enhancer and a constant (C) region comprising an endogenous CH gene segment; wherein said human variable region gene segments in said chimeric IgH locus are operably linked to said C region at a human/mouse chimeric junction within the JC intron of said chimeric IgH locus; wherein said homozygous chimeric IgH locus comprises in 5′ to 3′ transcriptional orientation:
(i) unrearranged human immunoglobulin heavy chain (IgH) variable region (VH) DNA comprising human IgH V gene segments, human D gene segments and human JH gene segments comprising a human 3′JH gene segment,
(ii) a chimeric J/C intron comprising human DNA downstream of and naturally contiguous with said human 3′ JH gene segment, which is contiguous with mouse JC intronic DNA, and
(iii) said enhancer and said C region,
wherein said human 3′JH is less than 1 kb upstream of said chimeric junction,
wherein DNA between said chimeric junction and said enhancer comprises mouse 129 strain JC intronic DNA;
wherein said enhancer is a mouse 129 strain μ enhancer;
wherein said transgenic mouse is functional to form rearranged human VH, DH and JH gene segments and to express chimeric immunoglobulin heavy chain polypeptide comprising a human VH region and a mouse C region, and wherein said transgenic mouse is capable, upon stimulation with antigen, of producing an antibody comprising a chimeric Ig heavy chain comprising a human IgH variable region and said C region; and
wherein the genome of said transgenic mouse comprises all or part of the endogenous mouse IgH variable region, and is capable of breeding to produce subsequent generation mice having in their germline an IgH locus comprising unrearranged human IgH variable region gene segments positioned upstream of an IgH constant (C) region comprising an endogenous C gene segment of an IgH locus,

(b) contacting said transgenic mouse with said selected antigen for a time and under conditions which permit the contacted mouse to produce antibody specific for said antigen; and
(c) isolating from said contacted mouse one or more of: antibody specific for said antigen, and/or a biological sample comprising antibody specific for said antigen, wherein the antibody comprises a chimeric immunoglobulin heavy chain polypeptide comprising a human VH region and a mouse C region.

2. The method of claim 1 , wherein said isolated biological sample comprises nucleic acid encoding said human VH region, and said method further comprises the step of:
isolating nucleic acid encoding said human VH region.

3. The method of claim 2 , further comprising the step of:
joining said human VH region encoding nucleic acid to a human constant region encoding nucleic acid, to provide nucleic acid encoding an antibody comprising said human VH region and a human constant region.

4. The method of claim 1 , wherein said method further comprises the step of: isolating said antibody from said isolated biological sample.

5. The method of claim 1 , wherein said unrearranged human heavy chain V region DNA comprises gene segments V6-1 to JH6 in human germline DNA order.

6. The method of claim 1 , wherein said said human VH region of said chimeric immunoglobulin heavy chain polypeptide is encoded by rearranged human variable region gene segments comprising a rearranged human VH2-5, human VDJH1, or human VDJH6 gene segment.

7. The method of claim 1 , wherein said human DNA contiguous with said human 3′JH gene segment comprises 400 base pairs of human JC intron DNA.

8. The method of claim 3 , further comprising the step of: expressing the antibody from said nucleic acid, wherein said nucleic acid encodes an antibody comprising said human variable region and a human constant region.

Assignees

• Kymab Ltd

Inventors

• Bradley Allan
• Lee E-Chiang
• Liang Qi
• Wang Wei
• Friedrich Glenn

Classifications

• A01K67/0275

  Genetically modified vertebrates, e.g. transgenic · CPC title

• C07K16/18 Primary

  against material from animals or humans · CPC title

• C07K16/1239

  from Vibrionaceae (G) · CPC title

• A61K39/107

  Vibrio · CPC title

• A01K2227/105

  Murine · CPC title