Non-human transgenic mammal expressing a human FcRn on its mammary gland cells and expressing a transgenic protein-human Fc-domain fusion

• US 8,173,860 B2
• Filed: 04/20/2007
• Issued: 05/08/2012
• Est. Priority Date: 04/21/2006
• Status: Expired due to Fees

• Current Assignee: LFB USA, Inc. (Agence des Participations de l’Etat)
• Original Assignee: GTC Biotherapeutics, Inc.
• Inventors: Meade, Harry M., Pollock, Daniel
• Primary Examiner(s): Crouch, Deborah
• Application Number:
  US11/788,775
• Publication Number:
  US 20080063780A1
• Time in Patent Office: 1,845 Days
• Field of Search: None
• US Class Current: 800/14
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: C07K 14/70535 Fc-receptors, e.g. CD16, CD32, CD64 (CD2314/705F)
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2510/02 Cells for production
• CPC Class Codes: C12N 2517/04 Cells produced using nuclear transfer
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination

Abstract

The invention provides, in part, methods for the production of proteins in a transgenic non-human mammal, wherein the proteins are transported from the blood to the mammary gland for secretion in milk. The transport of the protein to the mammary gland and/or milk is facilitated by binding to a transport receptor in the mammary gland.

Claims

1. A non-human transgenic mammal whose genome comprises a DNA sequence encoding human FcRn and a DNA sequence encoding beta-microglobulin operably linked to a mammary gland-specific promoter, and further comprises a DNA sequence encoding a transgenic protein comprising a human Fc-domain operably linked to a non-mammary gland-specific promoter.

2. The non-human mammal of claim 1, wherein the non-mammary gland-specific promoter is a liver-specific promoter.

3. The non-human mammal of claim 2, wherein the liver-specific promoter is an albumin promoter.

4. The non-human mammal of claim 1, wherein the transgenic protein is an antibody.

5. A composition comprising milk obtained from the non-human mammal of claim 1.

6. A method for obtaining a transgenic protein comprising a human Fc-domain, comprising:

generating a non-human transgenic mammal whose genome comprises a DNA sequence encoding human FcRn and a DNA sequence encoding beta-microglobulin operably linked to a mammary gland-specific promoter, and further comprises a DNA sequence encoding transgenic protein comprising a human Fc-domain operably linked to a non-mammary gland-specific promoter,

wherein the transgenic protein comprising a human Fc-domain binds the functional human FcRn via the human Fc-domain,

collecting milk from the non-human mammal, and

separating the transgenic protein comprising a human Fc-domain from the milk.

7. The method of claim 6, wherein the non-mammary gland-specific promoter is a liver-specific promoter.

8. The method of claim 7, wherein the liver-specific promoter is an albumin promoter.

9. The method of claim 6, wherein the transgenic protein comprising a human Fc-domain is an antibody.

10. A composition comprising the collected milk of the method of claim 6.

11. A composition comprising milk from a non-human transgenic mammal whose genome comprises a DNA sequence encoding human FcRn and a DNA sequence encoding beta-microglobulin operably linked to a mammary gland-specific promoter, and further comprises a DNA sequence encoding a transgenic protein comprising a human Fc-domain operably linked to a non-mammary gland-specific promoter.

12. The composition of claim 11, wherein the glycosylation pattern of the transgenic protein is different from the glycosylation pattern of the protein when expressed in the mammary gland of the non-human mammal.