PROCESS FOR PURIFYING INSULIN

• US 3,876,623 A
• Filed: 05/09/1973
• Issued: 04/08/1975
• Est. Priority Date: 05/09/1973
• Status: Expired due to Term

• Current Assignee: John G. Stilz, Richard L. Jackson
• Original Assignee: John G. Stilz, Richard L. Jackson
• Inventors: Jackson, Richard L., Stilz, John G.
• Primary Examiner(s): Gotts, Lewis
• Assistant Examiner(s): Suyat, Reginald J.
• Application Number:
  US05/358,554
• Time in Patent Office: 699 Days
• Field of Search: 260/112.5,112.7
• US Class Current: 530/305
• CPC Class Codes: C07K 14/625 Extraction from natural sources
• CPC Class Codes: Y10S 530/857 Fish; fish eggs; shell fish; crustacea

Abstract

Alkali metal or ammonium insulin is purified by gel filtration.

Claims

1. A PROCESS FOR PURIFYING AN ALKALI METAL OR AMMONIUM INSULIN WHICH COMPRISES THE STEPS OF:

A. SWELLING A GEL HAVING IN THE DRY STATE A WATER REGIN OF AT LEAST ABOUT 4 PERCENT BY WEIGHT AND PARTICLE DIAMETERS SMALLER THAN ABOUT 100 MICRONS, B. PACKING A COLUMN WITH THE SWOLLEN GEL, C. ADDING TO THE PACKED COLUMN AN AQUEOUS SOLUTION OF AN ALKALI METAL OR AMMONIUM INSULIN HAVING A PURITY OF AT LEAST ABOUT 80 PERCENT, WHEREIN THE INSULIN CONCENTRATION IS IN THE RANGE OF FROM ABOUT 1 TO ABOUT 8 PERCENT, WEIGHT PER VOLUME, THE TOTAL AMOUNT OF INSULIN IS SULFICIENT TO PROVIDE A COLUMN LOADING IN THE RANGE OF FROM ABOUT 0.8 TO ABOUT 6.7 GRAMS PER LITER OF BED VOLUME, AND THE PH OF THE SOLUTION IS IN THE RANGE OF FROM ABOUT 7.5 TO ABOUT 9.5, AND D. ELUTING THE INSULIN FROM THE COLUMN, AT A TEMPERATURE IN THE RANGE OF FROM ABOUT 5* TO ABOUT 30*C, WITH AN AQUEOUS ELUANT HAVING A PH IN THE SAME RANGE AS THAT OF THE INSULIN SOLUTION.

2. The process of claim 1, wherein said gel has a water regain in the range

3. The process of claim 2, wherein said gel has a water regain in the range

5. The process of claim 4, wherein said gel has a water regain of about 5

6. The process of claim 5, wherein the particle diameters of said gel are

7. The process of claim 1, wherein the column loading is in the range of

8. The process of claim 7, wherein the column loading is about 4.5 grams

9. The process of claim 1, wherein the eluant contains an inorganic salt at

10. The process of claim 9, wherein the inorganic salt is present at a

11. The process of claim 10, wherein said inorganic salt is sodium

12. The process of claim 1, wherein elution is carried out at ambient

13. The process of claim 1, wherein elution is carried out at 5.degree.C.