Genetic engineering of non-human animals for the production of chimeric antibodies
Inventors
Green, Larry • Shizuya, Hiroaki
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11220555-B2
Publication Date
2022-01-11
Expiration Date
Abstract
The invention provides non-human cells and mammals having a genome encoding chimeric antibodies and methods of producing transgenic cells and mammals. Certain aspects of the invention include chimeric antibodies, humanized antibodies, pharmaceutical compositions and kits. Certain aspects of the invention also relate to diagnostic and treatment methods using the antibodies of the invention.
Core Innovation
The invention provides a transgenic mouse whose genome comprises a transgene encoding a human immunoglobulin heavy chain variable region system. The transgene comprises, in operable linkage from 5′ to 3′, gene segments encoding a human variable heavy (VH) polypeptide, a human diversity heavy (DH) polypeptide, and a human joining heavy (JH) polypeptide.
The heavy-chain transgene is functionally linked to a constant region gene containing an exon encoding a rat CH1 polypeptide, an exon encoding a mouse CH2 polypeptide, and an exon encoding a mouse CH3 polypeptide. The constant region gene is operably linked with the gene segments encoding VH, DH, and JH, thereby providing a chimeric constant region configuration for the expressed heavy chain.
The document further describes genetically engineered non-human cells or transgenic mammals designed to produce chimeric or humanized antibodies by using engineered immunoglobulin heavy and light chain transgenes. It describes forcing antibody generation from engineered transgenes by optionally inactivating endogenous immunoglobulin loci, and it further includes concepts for designing and assembling transgenes so that the engineered systems support antibody expression and diversity, including somatic mutation and affinity maturation.
In the provided examples, the document describes synthetic lambda light-chain transgenes and transgenic mouse generation and expression, including integration confirmation and circulating human Igλ expression with repertoire diversity and somatic mutation. Overall, the invention addresses expression of chimeric/humanized antibodies in a transgenic animal context by combining engineered immunoglobulin gene segments with configured constant-region exons and, optionally, inactivation of endogenous Ig loci.
Claims Coverage
The independent claim covers a transgenic mouse with a heavy-chain transgene encoding human VH, DH, and JH gene segments operably linked to a chimeric constant region comprising rat CH1, mouse CH2, and mouse CH3 exons. The inventive features are further constrained in dependent claims by placement of constant-region exons, relationship to the endogenous IgH locus, functional inactivation of endogenous IgH, and optional addition and composition of a human light-chain transgene including Igλ elements.
Chimeric heavy-chain constant region with rat CH1 and mouse CH2/CH3
A mouse genome comprising a transgene with, in operable linkage, a constant region gene that includes an exon encoding a rat CH1 polypeptide, an exon encoding a mouse CH2 polypeptide, and an exon encoding a mouse CH3 polypeptide.
Operable 5′-to-3′ human VH, DH, and JH gene segments linked to the constant region gene
A mouse genome comprising a transgene where gene segments encoding a human variable heavy (VH) polypeptide, a human diversity heavy (DH) polypeptide, and a human joining heavy (JH) polypeptide are operably linked from 5′ to 3′ and are operably linked with the constant region gene.
Constant-region gene placement on an additional transgene
The mouse further has the constant region gene located on an additional transgene.
Relationship to the endogenous IgH locus via transgene replacement
The mouse has a transgene that replaces all or part of an endogenous IgH locus.
Functional inactivation of endogenous IgH
All or part of the mouse’s endogenous IgH locus is functionally inactivated.
Additional human λ light-chain transgene composition constraints
A second transgene containing human Vλ gene segments, human Jλ gene segments in the range of 1 to 7, and a single human Cλ gene segment.
Igλ 3′LCR obtained from a selected mammal group
The Igλ 3′LCR is obtained from a mammal selected from human, non-human primate, or rat.
Across the independent claim and dependent refinements, the core claim centers on operable linkage of human VH/DH/JH gene segments to a chimeric constant-region gene containing rat CH1 and mouse CH2/CH3 exons, with further constraints addressing constant-region placement, replacement/inactivation of endogenous IgH, and optional human λ light-chain transgene composition including source options for an Igλ 3′LCR.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
Interested in licensing this patent?