Methods of producing HBsAg

• US 4,416,986 A
• Filed: 01/16/1981
• Issued: 11/22/1983
• Est. Priority Date: 01/16/1981
• Status: Expired due to Term

• Current Assignee: Merck & Co., Inc.
• Original Assignee: Merck & Co., Inc.
• Inventors: Markus, Henry Z., McAleer, William J.
• Primary Examiner(s): Kepplinger, Esther M.
• Application Number:
  US06/225,555
• Time in Patent Office: 1,040 Days
• Field of Search: 435/68, 435/241, 435/948, 435/240
• US Class Current: 435/70.3
• CPC Class Codes: C07K 14/005 from viruses
• CPC Class Codes: C12N 2730/10122 New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Abstract

Hepatitis B surface antigen (HBsAg) is produced in vitro in high titer and purity from tissue cultures of cells that shed HBsAg. The cells are grown on hollow fiber capillary units having a molecular weight cut-off of about 10,000.

Claims

1. A method for preparing hepatitis B surface antigen which comprises growing cells which shed hepatitis B surface antigen in the presence of a nutrient medium on hollow fiber capillary units having a molecular weight cut-off of about 10,000.

2. A method according to claim 1 wherein the growing is effected with two sequential stages having differing temperatures, each temperature being above room temperature with the first temperature being above the second temperature.

3. A method according to claim 2 wherein the first temperature is from about 35°

to about 38°

C.

4. A method according to claim 2 wherein the second temperature is from about 30°

C. to about 34°

C.

5. A method according to claim 2 wherein the first temperature is about 37°

C. and the second temperature is about 32°

C.

6. A method according to claim 2 wherein caffeine is present during the second stage.

7. A method according to claim 6 wherein the caffeine is present in an amount from about 0.0001 M to about 0.0003 M.