Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Histidine Engineered Light Chain Antibodies and Genetically Modified Non-Human Animals for Generating the Same

a technology of light chain antibody and histidine, which is applied in the field of histidine engineered light chain antibody and genetically modified non-human animals for generating the same, can solve the problems of bispecific antibody having a suitable light chain component that can satisfactorily associate with each of the heavy chains of the bispecific antibody, requiring high doses to achieve the desired effect, and reducing binding

Inactive Publication Date: 2014-01-09
REGENERON PHARM INC
View PDF4 Cites 49 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes a genetically modified non-human animal that produces antibodies that have a reduced binding to a target antigen at an acidic pH compared to a neutral pH. This is achieved by modifying the light chain sequence of the animal to include one or more histidine modifications in the light chain CDR3 codon. The non-human animal also has a replacement of endogenous non-human heavy chain constant region gene sequences with human sequences. The technical effect of this modification is that the animal produces antibodies that have a decreased dissociative half-life at an acidic pH compared to a neutral pH.

Problems solved by technology

But making bispecific antibodies having a suitable light chain component that can satisfactorily associate with each of the heavy chains of a bispecific antibody has proved problematic.
In addition, therapeutic antibodies, e.g., bispecific therapeutic antibodies, have some limitations in that they often require high doses to achieve desired efficacy.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Histidine Engineered Light Chain Antibodies and Genetically Modified Non-Human Animals for Generating the Same
  • Histidine Engineered Light Chain Antibodies and Genetically Modified Non-Human Animals for Generating the Same
  • Histidine Engineered Light Chain Antibodies and Genetically Modified Non-Human Animals for Generating the Same

Examples

Experimental program
Comparison scheme
Effect test

example 1

Identification of Histidine Residues in Antigen-Specific Human Light Chains

[0258]Generation of a common light chain mouse (e.g., Vκ1-39 or Vκ3-20 common light chain mouse) and antigen-specific antibodies in those mice is described in, e.g., U.S. patent application Ser. Nos. 13 / 022,759, 13 / 093,156, and 13 / 412,936 (Publication Nos. 2011 / 0195454, 2012 / 0021409, and 2012 / 0192300, respectively), incorporated by reference herein in their entireties. Briefly, rearranged human germline light chain targeting vector was made using VELOCIGENE® technology (see, e.g., U.S. Pat. No. 6,586,251 and Valenzuela et al. (2003) High-throughput engineering of the mouse genome coupled with high-resolution expression analysis, Nature Biotech. 21(6): 652-659) to modify mouse genomic Bacterial Artificial Chromosome (BAC) clones, and genomic constructs were engineered to contain a single rearranged human germline light chain region and inserted into an endogenous κ light chain locus that was previously modifie...

example 2

Engineering and Characterization of Histidine-Substituted Human Universal Light Chain Antibodies

example 2.1

Engineering of Histidine Residues into a Germline Human Rearranged Light Chain

[0261]Histidine residues were engineered into a rearranged human Vκ1-39Jκ5 light chain using site directed mutagenesis primers specifically designed to introduce engineered histidine residues at Q105, Q106, Y108, and P111 positions of the human Vκ1-39Jκ5 light chain. Site directed mutagenesis was performed using molecular techniques known in the art (e.g., QuikChange II XL Site Directed Mutagenesis Kit, Agilent Technologies). Locations of the engineered residues in the CDR3 are shown in FIG. 2, the nucleic acid sequences of histidine-substituted CDR3's depicted in FIG. 2 are set forth in SEQ ID NOs: 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, and 32 (corresponding amino acid sequences are set forth in SEQ ID NOs: 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, and 33). The nucleic acid and amino acid sequences of germline rearranged Vκ1-39Jκ5 CDR3 are set forth in SEQ ID NOs: 2 and 3, respect...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Lightaaaaaaaaaa
Affinityaaaaaaaaaa
Login to View More

Abstract

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 human immunoglobulin light chain variable domains derived from a limited repertoire of human immunoglobulin light chain variable gene segments that comprise histidine modifications in their germline sequence. Methods of making non-human animals that express antibodies comprising histidine residues encoded by histidine codons introduced into immunoglobulin light chain nucleotide sequences are provided.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation-in-part of U.S. patent application Ser. No. 13 / 832,247, filed Mar. 15, 2013, which claims benefit under 35 U.S.C. §119(e) of U.S. Provisional Application No. 61 / 611,950, filed Mar. 16, 2012, and U.S. Provisional Application No. 61 / 736,930, filed Dec. 13, 2012, all incorporated by reference herein in their entireties.FIELD OF INVENTION[0002]A genetically modified non-human animal (e.g., rodent, e.g., mouse or rat) is provided that expresses antibodies capable of binding to an antigen in a pH dependent manner. A method for making modifications to immunoglobulin light chain variable region sequence of a non-human animal is provided, wherein the modifications include the mutagenesis of residues within the light chain variable region gene, e.g., nucleotides that encode one or more amino acids within a complementary determining region (CDR), to facilitate in vivo expression of antibodies comprising light chain...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K16/00C12N15/85A01K67/027
CPCC07K16/00A01K67/0275C12N15/8509A01K67/0278A01K2217/072A01K2217/075A01K2217/15A01K2227/105A01K2267/01C07K2317/515C07K2317/92C12N2800/204C07K2317/21C07K2317/94C07K2317/51C07K2317/24C07K2317/56
Inventor MCWHIRTER, JOHNMACDONALD, LYNNMURPHY, ANDREW J.
Owner REGENERON PHARM INC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com