Murine expression of a human IgA lambda locus

• US 20020088016A1
• Filed: 12/13/2000
• Published: 07/04/2002
• Est. Priority Date: 11/03/1998
• Status: Active Grant

• Current Assignee: The Babraham Institute
• Original Assignee: The Babraham Institute
• Inventors: Bruggemann, Marianne
• Granted Patent: US 6,998,514 B2
• Time in Patent Office: Days
• US Class Current: 800/18
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/072 maintaining or altering function, i.e. knock in
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 2267/0381 Animal model for diseases of the hematopoietic system
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic

Abstract

In humans, approximately 60% of expressed immunoglobulin light chains are of the Kappa type and 40% of the Lambda type. In mice, there is almost no expression from the Lambda locus and over 95% of light chains are of Kappa type. The present invention discloses, among other things, transgenic mice carrying most of the human Ig Lambda light chain locus in their genome. The resulting mice express light chains with Kappa/Lambda ratio similar to the human ratio. Breeding of HuIg Lamda mice to Kappa-deficient mice also is described, as well as the generation of human monoclonal antibodies from transgenic mice with human Ig Lambda locus.

Claims

1. A transgenic mouse comprising as a translocus a YAC of about 410 Kb, wherein the YAC contains most of the human Vλ

genes of cluster A and all the human Jλ

-Cλ

segments in germline configuration, wherein the translocus shows high expression, and is able to compete equally with the endogenous mouse κ

locus.

5. A transgenic mouse according to claim 1, wherein the mouse includes a HuIgλ

YAC that accommodates a 380 Kb region of the human λ

light chain locus in authentic configuration with all Vλ

genes of cluster A, the Jλ

-Cλ

segments and the 3′

enhancer.

6. A transgenic mouse according to claim 5, wherein the HuIgλ

YAC is shown in FIG. 1.

7. A method for producing a transgenic mouse according to claim 1, comprising:

(a) introducing a HuIgλ

YAC into murine embryonic stems cells; and

(b) deriving a transgenic mouse from the cells of step (a).

8. The method of claim 7, wherein a HuIgλ

YAC of about 410 Kb that can accommodate a 380 Kb region (Vλ

-JCλ

) of the human λ

light chain locus with V, J and C genes in germline configuration is introduced into said stem cells.

9. The method according to claim 7 wherein two copies of the neomycin resistance gene (NBO^r) are site-specifically integrated into the ampicillin gene on the left (centromeric) YAC arm in order to permit selection.

2. A transgenic mouse comprising as a translocus a YAC of about 410 Kb, wherein the YAC contains most of the human Vλ

genes of cluster A and all the human Jλ

-Cλ

segments in germline configuration, wherein the mouse has one or both endogenous Igκ

alleles disrupted, and wherein the translocus shows high expression.

3. A transgenic mouse carrying a 380 Kb region of the human immunoglobulin (Ig) λ

light (L) chain locus in germline configuration, wherein the introduced translocus resides on a yeast artificial chromosome (YAC) that accommodates the most proximal V (variable gene) λ

cluster—

with 15 V λ

genes that contribute to over 60% of λ

light chains in man—

and all Jλ

-Cλ

segments with the 3′

region including the downstream enhancer.

4. A transgenic mouse comprising human Ig lambda genes in which the proportion of the κ

and λ

light chains expressed by said human lambda genes resembles that found in humans, and exhibits relative proportions of ≦

60% κ

light chains and ≧

40% λ

light chains.