Production of membrane proteins in the milk of transgenic mammals

• US 20030033618A1
• Filed: 06/27/2002
• Published: 02/13/2003
• Est. Priority Date: 10/02/1991
• Status: Active Grant

• Current Assignee: Genzyme Corporation (Sanofi )
• Original Assignee: Genzyme Corporation (Sanofi )
• Inventors: Smith, Alan E.
• Granted Patent: US 6,743,966 B2
• Time in Patent Office: Days
• 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 2227/10 Mammal
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/01 Animal expressing industrially exogenous proteins
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic

Abstract

Production of proteins not normally secreted through conventional pathways such as membrane proteins including, for example, CFTR associated with cystic fibrosis, is now made possible by collection of such protein from the milk of lactating transgenic animals.

Claims

1. A method for producing a membrane protein comprising the steps of:

(a) inserting into a mammalian embryo a DNA construct comprising a mammary gland specific promoter and a DNA sequence encoding said membrane protein,

(b) allowing said embryo to mature into a reproductively competent mammal,

(c) inducing lactation in females of said reproductively competent mammal or in female progeny of said reproductively competent mammal, and

(d) collecting the milk of said lactating female which contains said membrane protein.

2. The method of claim 1 wherein said mammary gland specific promoter is selected from the group of genes consisting of betalactoglobulin, alpha casein, beta casein and whey acid protein encoding genes.

3. The method of claim 2 wherein said membrane protein is CFTR.

4. The method of claim 3 further comprising the step of isolating the CFTR from the collected milk.

5. The method of claim 2 wherein said membrane protein is selected from the group consisting of hormone receptor proteins, cytokine receptor proteins, viral glycoproteins and transmembrane channels.

6. The mammalian embryo produced by the method of claim 1.

7. The mammal produced by the method of claim 1.

8. The membrane protein produced by the method of claim 1.

9. A method for producing a membrane protein comprising collecting milk comprising said membrane protein from a transgenic mammal having in its genome a gene encoding said membrane protein and under the control of a mammary gland specific protein promoter.

10. The method of claim 9 wherein said mammary gland specific promoter is selected from the group of genes consisting of betalactoglobulin, alpha casein, beta casein and whey acid protein encoding genes.

11. The method of claim 10 wherein said membrane protein is selected from the group consisting of hormone receptor proteins, cytokine receptor proteins, viral glycoproteins and transmembrane channels.

12. The method of claim 10 wherein said membrane protein is CFTR.

13. The method of claim 12 further comprising the step of isolating the CFTR from the collected milk.

14. A method for producing a membrane protein comprising culturing cells transformed with a gene encoding a membrane protein and capable of apocrine secretion and collecting said membrane protein from the cell culture media.

15. The method of claim 14 wherein said membrane protein is selected from the group consisting of hormone receptor proteins, cytokine receptor proteins and transmembrane channels.