Transgenic ungulates capable of human antibody production

• US 7,803,981 B2
• Filed: 06/20/2007
• Issued: 09/28/2010
• Est. Priority Date: 11/19/1999
• Status: Expired due to Term

• Current Assignee: SAB LLC (W.P. Carey, Inc. (Real Estate))
• Original Assignee: Kyowa Hakko Kirin Company Limited (Kirin Holdings Company Limited)
• Inventors: Osborne, Barbara, Sullivan, Eddie, Collas, Philippe, Kuroiwa, Yoshimi, Goldsby, Richard A., Tomizuka, Kazuma, Ishida, Isao, Robl, James M., Kasinathan, Poothappillai
• Primary Examiner(s): Crouch; Deborah
• Application Number:
  US11/820,768
• Publication Number:
  US 20080040821A1
• Time in Patent Office: 1,196 Days
• Field of Search: None
• US Class Current: 800/15
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0273 Cloned vertebrates
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• 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 16/42 against immunoglobulins
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 15/8771 Bovine embryos
• CPC Class Codes: C12N 2799/025 where the vector is derived from a parvovirus
• CPC Class Codes: C12N 2800/208 of mammalian origin, e.g. minichromosome

Abstract

The invention features novel methods for the production of large quantities of xenogenous antibodies, such as human antibodies. Preferably, this result is effected by inactivation of IgM heavy chain expression and, optionally, by inactivation of Ig light chain expression, and by the further introduction of an artificial chromosome which results in the expression of xenogenous antibodies (e.g., non-bovine antibodies), preferably human antibodies.

Claims

1. A transgenic ungulate whose genome comprises a mutation in an endogenous immunoglobulin (Ig) heavy chain or Ig light chain locus, said mutation reducing the expression of said Ig heavy chain or Ig light chain.

2. The ungulate of claim 1, wherein said mutation reduces the expression of functional IgM heavy chain.

3. The ungulate of claim 1, wherein said mutation substantially eliminates the expression of said Ig heavy chain or said Ig light chain.

4. The ungulate of claim 1, wherein said mutation is hemizygous.

5. The ungulate of claim 1, wherein said mutation is homozygous.

6. The ungulate of claim 1, wherein said mutation is an insertion of a positive selection marker into said endogenous locus.

7. The ungulate of claim 6, wherein said positive selection marker is an antibiotic resistance gene.

8. The ungulate of claim 7, wherein each allele comprises the same antibiotic resistance gene.

9. The ungulate of claim 7, wherein each allele comprises a different antibiotic resistance gene.

10. The ungulate of claim 6, wherein said positive selection marker is operably linked to a xenogenous promoter.

11. The ungulate of claim 1, wherein said mutation is an insertion of a transcription termination sequence into said endogenous loci.

12. The ungulate of claim 11, wherein said transcription termination sequence is inserted downstream of the initial ATG codon in exon 2 of an endogenous mu heavy chain locus.

13. The ungulate of claim 1, comprising one or more nucleic acids comprising one or more transgenes and expressing an mRNA or protein encoded by said transgene(s).

14. The ungulate of claim 1, wherein said ungulate is a pig.

15. An isolated ungulate somatic cell comprising a mutation in an endogenous immunoglobulin (Ig) heavy chain or Ig light chain locus, said mutation reducing the expression of said Ig heavy chain or Ig light chain.

16. The cell of claim 15, comprising a mutation in both alleles of said IgM heavy chain or said light chain.

17. The cell of claim 15, wherein said mutation is a transcription termination sequence.

18. The cell of claim 17, wherein said transcription termination sequence is inserted downstream of the initial ATG codon in exon 2 of an endogenous mu heavy chain locus.

19. The cell of claim 15, wherein said cell is a fetal fibroblast or a B-cell.

20. The cell of claim 15, wherein said ungulate is a pig.