Genetic engineering of non-human animals for the production of chimeric antibodies

Inventors

Green, Larry • Shizuya, Hiroaki

Assignees

Ablexis LLC

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 platform for engineering non-human mammalian cells and non-human animals to produce chimeric and/or humanized antibodies. It centers on constructing immunoglobulin loci in which chimeric immunoglobulin heavy chain variable (V) genes are combined with non-endogenous immunoglobulin constant region elements, including a non-endogenous CH1 domain, and optionally engineered Ig light chain elements.

A key problem addressed is that existing transgenic/knock-in and antibody generation methods have drawbacks, and that antibody generation systems require proper endogenous control so that V(D)J recombination, class switching, and B-cell development occur correctly. The disclosure describes preserving functional cis regulatory elements between immunoglobulin V gene segments and adjacent regulatory regions, including promoters and recombination signal sequences (RSS), as well as locus control region (LCR) and enhancers, to support correct recombination processes, class switch recombination (CSR), and B-cell development.

The invention also describes tailored immunoglobulin loci constructed using in silico sequence design and synthetic DNA assembly, including BAC-based assemblies. It further describes strategies to inactivate endogenous Ig loci so that the synthetic or engineered immunoglobulin loci function without interference from native loci, and reports synthetic Ig lambda (Lambda-Prime, Lambda 3/5) and recombined/engineered Ig kappa transgenes that lead to transgenic mice expressing diverse human Ig light chains with somatic hypermutation and affinity maturation.

Claims Coverage

The partial claim set provided includes one independent claim. The inventive features focus on a murine cell transgene architecture that positions human Ig heavy-chain V genes with mouse/rat immunoglobulin non-coding and mouse/rat cis regulatory sequences, with additional refinement options for regulatory element types and adding light-chain transgene components.

Across the provided partial independent-claim text, the core claim elements are the murine cell transgene that combines human immunoglobulin heavy chain variable (V) genes with mouse/rat immunoglobulin non-coding sequences and mouse/rat cis regulatory sequences positioned between the V genes. The additional refinements shown in the provided dependents further narrow the transgene characterization as synthetic and specify that the cis regulatory sequences are mouse, rat, or a combination thereof.

Stated Advantages

Documented Applications