IP Library Granted Patent US 9,976,163
Granted Patent B2
US 9,976,163 · App. 14/432,958 · Granted May 22, 2018

Method for preventing reduction of polypeptide by adding amino acid to culture solution

Inventor: Hidetoshi Inoue (Tokyo, JP)
Assignee: KYOWA HAKKO KIRIN CO., LTD
C12P21/00C07K16/00C12P21/02C07K2317/14
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Quick Facts
Patent No.
US 9,976,163
App. No.
14/432,958
Granted
May 22, 2018
Kind
B2
Abstract

The present invention provides a method for preventing the reduction of a desired polypeptide during purification in order to purify the polypeptide in high yield from a culture solution which contains recombinant host cells and the polypeptide.

Claims (15)

1. A method for purifying a polypeptide from a culture solution containing the polypeptide and a recombinant host cell which is capable of producing the polypeptide, said method comprising the following steps:

(a) a flocculation step, wherein the flocculation step comprises acid precipitation of the culture solution;

(b) centrifuging the culture solution of step (a) to recover a culture supernatant;

(c) filtering the culture supernatant; and

(d) isolating the polypeptide from the filtrate obtained from step (c),

wherein the method further comprises adding arginine to the culture solution either before or after step (a).

2. The method according to claim 1 , wherein the polypeptide is an antibody or an antibody fragment.

3. The method according to claim 1 , wherein the recombinant host cells are Chinese hamster ovary (CHO) cells.

4. The method according to claim 1 , wherein the arginine is added to the culture solution before step (a).

5. The method according to claim 1 , wherein the arginine is added to the culture solution after step (a).

6. The method according to claim 5 , wherein the arginine is added to the culture solution so that the final concentration of the arginine is from 0.05 mol/L to 1 mol/L.

7. The method according to claim 5 , wherein the arginine is added to the culture solution so that the final concentration of the arginine is 0.5 mol/L.

8. The method according to claim 5 , wherein the arginine is added to the culture solution so that the final concentration of the arginine is 0.1 mol/L.

9. The method according to claim 5 , further comprising adding arginine to the culture solution after step (c).

10. The method according to claim 5 , wherein arginine is added to the culture solution after step (a), and before step (b).

Assignments (2)
CHANGE OF NAME AND ADDRESS Recorded Oct 23, 2019
From: KYOWA HAKKO KIRIN CO., LTD.
To: KYOWA KIRIN CO., LTD.
Reel/Frame 050797/0533 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 2, 2015
From: INOUE, HIDETOSHI
To: KYOWA HAKKO KIRIN CO., LTD.
Reel/Frame 035320/0449 →
Continuity (2)
Provisional Application 61709311 · Oct 3, 2012
Related Publication 20150275259A1 · Oct 1, 2015