Isolation of exogenous recombinant proteins from the milk of transgenic mammals

• US 4,873,316 A
• Filed: 06/23/1987
• Issued: 10/10/1989
• Est. Priority Date: 06/23/1987
• Status: Expired due to Term

• Current Assignee: Pharming Intellectual Property B.V. (Pharming Group N.V.)
• Original Assignee: Biogen, Inc.
• Inventors: Lonberg, Nils, Meade, Harry
• Primary Examiner(s): Moskowitz, Margaret
• Assistant Examiner(s): Kushan, Jeff P.
• Application Number:
  US07/065,994
• Time in Patent Office: 840 Days
• Field of Search: 435/68, 435/172.1, 435/172.3, 435/226, 435/240.2, 530/832, 530/833, 530/412, 530/360, 530/361, 530/303, 800/1, 935/53, 935/55, 935/70, 536/27, 536/28, 536/29
• 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 2227/103 Ovine
• 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: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: C07K 14/4732 Casein in foodstuffs A23J
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2830/008 cell type or tissue specific enhancer/promoter combination
• CPC Class Codes: C12N 2830/85 mammalian
• CPC Class Codes: C12N 2840/44 being a specific part of the splice mechanism, e.g. donor, acceptor
• CPC Class Codes: C12N 9/6456 Plasminogen activators
• CPC Class Codes: Y10S 530/832 Milk; colostrum
• CPC Class Codes: Y10S 530/833 Whey; cheese

Abstract

This invention relates to the production of recombinant proteins in mammals'"'"' milk. Particularly, this invention relates to an expression system comprising the mammal'"'"'s casein promoter which when transgenically incorporated into a mammal permits the female species of that mammal to produce the desired recombinant protein in or along with its milk. This invention also relates to the transgenic mammal that produces the desired recombinant product in its milk.

Claims

1. A process for the production and secretion into mammal'"'"'s milk of an exogenous recombinate protein comprising the steps of:

a. producing milk in a transgenic mammal characterized by an expression system comprising a casein promoter operatively linked to an exogenous DNA sequence coding for the recombinant protein through a DNA sequence coding for a signal peptide effective in secreting and maturing the recombinant protein in mammary tissue;

b. collecting the milk; and

c. isolating the exogenous recombinant protein from the milk.

2. The process according to claim 1, wherein said expression system also includes a 3'"'"' untranslated region downstream of the DNA sequence coding for the recombinant protein.

3. The process according to claim 1, wherein said expression system also includes a 5'"'"' untranslated region between said promoter and the DNA sequence coding for the signal peptide.