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 document describes genetically engineered non-human animals and transgenic cells for producing chimeric and humanized antibodies by providing immunoglobulin loci in which endogenous immunoglobulin components are replaced with non-endogenous immunoglobulin variable and constant region segments. In particular, it focuses on transgenes that replace immunoglobulin light chain loci such that human immunoglobulin light chain variable (V) regions and human immunoglobulin light chain joining (J) regions are encoded together with configured mouse/rat immunoglobulin light chain non-coding sequences between V and J regions and between J-region elements.

A central aspect is the use of mouse or rat immunoglobulin non-coding sequences located between the light chain V exons and between the J coding sequences to support the configured light-chain locus architecture. The document further specifies, in light-chain transgenes, arrangements that include multiple human light chain V exons, multiple human light chain J exons/coding sequences, and configured human/mouse pairings of J and C sequences within the transgene, with additional non-coding sequence content positioned between J–C coding sequence pairs.

The document also explains that light-chain transgene constructs can include cis regulatory and non-coding element configurations, with the stated rationale of preserving immunoglobulin domain conformation through engineering of relevant domain context and replacement or tailoring of constant-region and hinge sequences in the immunoglobulin architecture. It discusses the construction of large Igκ/Igλ locus BACs and synthetic Igλ light-chain transgenes and describes generating transgenic mice with intact integration and expression of diverse human light chains, with evidence of somatic mutation/affinity maturation, and further embodiments involving inactivation of endogenous Ig loci.

Claims Coverage

The partial set contains three independent claims directed to polynucleotide transgenes encoding human immunoglobulin light chains with mouse/rat non-coding sequences and defined human/mouse V, J, and C arrangements. Across the independent claims, the core coverage is implemented through inventive features that (i) specify the human light-chain V exons and human J coding sequences, (ii) place mouse/rat non-coding sequences between defined gene segments, and (iii) define which J and C sequences are human versus mouse within J–C coding sequence pairs and associated spacing.

Across the independent claims, the document claims polynucleotide transgenes that encode human immunoglobulin light chain variable V regions and human light-chain joining J regions, while positioning mouse/rat immunoglobulin non-coding sequences between V exons and between J coding sequences. The independent claim set further specifies human-versus-mouse pairing of J and constant C sequences (human Jλ with mouse Cλ) in Jλ–Cλ coding sequence pairs and, for κ, a mouse Cκ constant coding sequence, with additional dependent refinements constraining numbers of J–C pairs and non-coding regions at defined positions.

Stated Advantages

Documented Applications