Means and methods for raising antibody concentration in compartments of the body of a non-human animal

• US 6,924,412 B1
• Filed: 07/21/2000
• Issued: 08/02/2005
• Est. Priority Date: 07/21/2000
• Status: Expired due to Fees

• Current Assignee: Lactive B.V.
• Original Assignee: Arriwan Holding B.V.
• Inventors: de Boer, Herman Albert, de Groot, Nanda
• Primary Examiner(s): Priebe, Scott D.
• Assistant Examiner(s): WHITEMAN, BRIAN A
• Application Number:
  US09/621,593
• Time in Patent Office: 1,838 Days
• Field of Search: 800/4, 800/7, 800/8, 800/13, 800/14, 800/18, 800/25, 800/15, 800/10, 800/24, 424/93.21, 424/93.2, 435/455, 514/44
• US Class Current: 800/7
• 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 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 2267/02 Animal zootechnically ameliorated
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: C07K 14/70535 Fc-receptors, e.g. CD16, CD32, CD64 (CD2314/705F)
• 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 16/1239 from Vibrionaceae (G)
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic

Abstract

Methods and processes for raising the concentration of a first class of immunoglobulin relative to at least a second class of immunoglobulin in a compartment of the body of a non-human animal or the progeny thereof, as well as the animals produced by such methods and processes. Such methods and processes provide for the collection of antibodies produced by mucosal surfaces of the animal. Preferably, the production is in the mammary gland. Antibodies can be collected from the milk of the animal. Antibodies may be used for medical and/or nutritional purposes.

Claims

1. A transgenic, non-human mammalian animal whose genome comprises:

a stably integrated, recombinant nucleic acid encoding a polymeric immunoglobulin receptor (pIgR) protein operatively linked to a mammary specific promoter;

wherein a mammary gland of said transgenic, non-human mammalian animal over-expresses said pIgR protein as compared to expression of the pIgR protein in a mammary gland of a wild-type, non-human mammalian animal;

wherein said pIgR protein transports a polymeric immunoglobulin protein across the basolateral side of a mammary gland'"'"'s epithelial cell to the apical side of the mammary gland'"'"'s epithelial cell as compared to a different immunoglobulin protein located on the epithelial cell'"'"'s basolateral side.

2. The transgenic, non-human mammalian animal of claim 1, wherein the polymeric immunoglobulin protein is selected from the group consisting of IgM and IgA.

3. The transgenic, non-human mammalian animal of claim 1, wherein the immunoglobulin protein located on the epithelial cell'"'"'s basolateral side is IgG.

4. The transgenic, non-human mammalian animal of claim 1, wherein said transgenic, non-human mammalian animal over-expresses said pIgR protein at least 10-fold higher than the expression of the pIgR protein in the wild-type, non-human mammalian animal.

5. A method of making the transgenic, non-human mammalian animal of claim 1, said method comprising:

producing a DNA construct comprising a nucleic acid encoding the pIgR protein operably linked to a mammary specific promoter that drives expression of said pIgR protein in a mammary gland epithelial cell;

introducing said DNA construct into fertilized eggs; and

implanting the fertilized eggs into a pseudopregnant, female non-human mammalian animal, thereby producing the transgenic, non-human mammalian animal of claim 1.

6. The method according to claim 5, wherein said mammary specific promoter is a casein promoter.

7. A method of collecting an immunoglobulin from the transgenic, non-human mammalian animal of claim 1, comprising:

providing the transgenic, non-human mammalian animal of claim 1; and

collecting milk comprising said polymeric immunoglobulin protein from the mammary gland of said transgenic, non-human mammalian animal.

8. The method according to claim 7, further comprising isolating said polymeric immunoglobulin protein from the milk.

9. The method according to claim 7, wherein collecting milk comprising said polymeric immunoglobulin protein comprises collecting milk comprising IgM or IgA.

10. The method according to claim 7, further comprising administering an antigen to said transgenic, non-human mammalian animal prior to collecting the milk from the mammary gland.