Isolation of components of interest from milk

• US 5,756,687 A
• Filed: 01/23/1995
• Issued: 05/26/1998
• Est. Priority Date: 03/09/1993
• Status: Expired due to Term

• Current Assignee: Genzyme Transgenics Corporation
• Original Assignee: Genzyme Transgenics Corporation
• Inventors: Denman, Julie S., Cole, Edward S.
• Primary Examiner(s): Schwadron, Ronald B.
• Application Number:
  US08/376,620
• Time in Patent Office: 1,219 Days
• Field of Search: 435/69.1, 435/70.1, 435/71.1, 435/183, 435/226, 530/412, 530/413, 530/414, 530/350, 530/415, 530/416, 530/417, 530/418, 530/422, 530/832, 530/419
• US Class Current: 530/412
• 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 2227/102 Caprine
• 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: A23C 3/08 by addition of preservatives additions of microorganisms or enzymes A23C9/12, of other substances A23C9/152
• CPC Class Codes: A23J 1/20 from milk, e.g. casein curds or cheese A23C; from whey
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: Y10S 530/832 Milk; colostrum

Abstract

Methods of isolating components of interest from a milk sample are described. The methods include a step wherein the solubility of at least a portion of the total milk protein is stabilized in such a manner as to allow isolation of the component of interest without significant loss in yield. Kits for stabilizing the solubility of at least a portion of the total milk protein of the milk sample containing the component of interest also are described.

Claims

1. A method of isolating a protein of intrest from a milk sample, comprising:

contacting a milk sample containing a protein of interest with a cationic solubilizing agent under conditions which stabilize the solubility of at least a portion of the total milk protein such that the protein of interest can be isolated from the solubilized milk sample with less than 50% loss of the protein of interest; and

isolating the protein of interest from the solubilized milk sample in a form which is biologically active.

2. The method of claim 1 wherein the milk sample is obtained from a transgenic mammal.

3. The method of claim 1 wherein the protein of interest is a naturally occurring milk protein.

4. The method of claim 1 wherein the protein of interest is a protein secreted in the milk of a transgenic mammal.

5. The method of claim 1 wherein the protein of interest is a therapeutic agent.

6. The method of claim 5 wherein the therapeutic agent is longer-acting tissue-type plasminogen activator.

7. The method of claim 5 wherein the therapeutic agent is anti-thrombin III.

8. The method of claim 1 wherein the protein of interest is an immunoglobulin.

9. The method of claim 1 wherein the pH of the milk sample containing the cationic solubilizing agent ranges from about 5.0 to about 9.0.

10. The method of claim 1 wherein the catonic solubilizing agent is arginine.

11. The method of claim 1 wherein the cationic solubilizing agent is imidazole.

12. The method of claim 1 wherein the cantonic solubilizing agent is Bis-Tris.

13. The method of claim 1 further comprising filtering of the milk sample and isolating the protein of interest from the filtrate.

14. The method of claim 1 wherein the protein of intrest is isolated from the filtrate by chromatography.

15. The method of claim 1 wherein the loss of the protein of interest is less than about 20%.

16. The method of claim 1 wherein the loss of the protein of interest is less than about 10%.