Methods of producing a target molecule in a transgenic animal and purification of the target molecule

• US 20040226053A1
• Filed: 06/08/2004
• Published: 11/11/2004
• Est. Priority Date: 10/13/2000
• Status: Abandoned Application

• Current Assignee: Harry M. Meade, Scott Fulton, Yann Echelard
• Original Assignee: Harry M. Meade, Scott Fulton, Yann Echelard
• Inventors: Fulton, Scott, Echelard, Yann, Meade, Harry M.
• Application Number:
  US10/863,101
• Publication Number:
  US 20040226053A1
• Time in Patent Office: Days
• US Class Current: 800/7
• CPC Class Codes: C07K 1/22 Affinity chromatography or related techniques based upon selective absorption processes
• CPC Class Codes: C07K 1/36 by a combination of two or more processes of different types

Abstract

The invention provides systems and methods for the production and purification of target molecules present in biological systems. The systems and methods according to the invention utilize transgenic expression of multivalent binding polypeptides, as affinity media, to purify such target molecules. The transgenic multivalent binding polypeptides bind both the target molecules, e.g., a bindable epitope of a target molecule, and a matrix.

Claims

1-4. -4. Cancel

5. A method of obtaining a target polypeptide having a bindable epitope from a product, the method comprising:

contacting a product which comprises a target polypeptide having a bindable epitope with a transgenically produced multivalent binding polypeptide, wherein the transgenically produced multivalent binding polypeptide comprises a first binding moiety which specifically binds the bindable epitope of the target polypeptide and a second binding moiety which specifically binds a matrix, to thereby provide a reaction mixture;

contacting the reaction mixture with a matrix which specifically binds the second binding moiety of the multivalent binding polypeptide;

removing reaction mixture which does not bind to the matrix, to thereby obtain the target polypeptide from the product; and

wherein the reaction mixture is substantially fluid.

31. The method of claim 5, wherein said multivalent polypeptide is used in an ELISA format.

33. The method of claim 5, wherein said reaction mixture is semi-solid

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12. A method of obtaining a target polypeptide having a bindable epitope from the milk of a first non-human transgenic mammal, the method comprising:

contacting milk which comprises a target polypeptide having a bindable epitope with a transgenically produced multivalent binding polypeptide, wherein the multivalent binding polypeptide comprises a first binding moiety which specifically binds the bindable epitope of the target polypeptide and a second binding moiety which specifically binds a matrix, to thereby provide a reaction mixture;

contacting the reaction mixture with a matrix which specifically binds the second binding moiety of the multivalent binding polypeptide;

removing reaction mixture which does not bind to the matrix, to thereby obtain the target polypeptide from the milk;

wherein the reaction mixture is substantially fluid; and

,

wherein the transgenically produced multivalent binding polypeptide is produced in milk from a second non-human transgenic mammal.

34. The method of claim 12, wherein said multivalent polypeptide is used in an ELISA format.

35. The method of claim 12, wherein said target polypeptide is purified from the reaction mixture to a composition that is more than 90% pure.

36. The method of claim 12, wherein said reaction mixture is semi-solid.

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