Claims
- 1. A method for purifying a crude tPA preparation, containing tPA and undesired proteins, said method comprising the successive steps of:
- (a) contacting said crude preparation with a cation-exchanger having a carboxymethyl group as the cation-exchange group to adsorb the APA and proteins onto said cation-exchanger;
- (b) washing said cation-exchanger with an eluent having a pH in the range between 5.2 and 6.5 at a salt concentration at which the tPA is not eluted to elute the undesired proteins having PI values equivalent to or lower than that of said tPA;
- (c) washing said cation-exchanger with an eluent having a pH in the range between 2.8 and 3.5 at a salt concentration at which the tPA is not eluted to elute the undesired proteins having the PI values equivalent to or higher than that of said tPA: and
- (d) eluting tPA having a molecular weight of about 70,000 daltons and a PI value in the range 6 to 8 from the cation-exchanger at a pH of below 2.8.
- 2. The method of claim 1, wherein the crude tPA preparation has a pH in the range 4.5 to 5.0
Priority Claims (1)
Number |
Date |
Country |
Kind |
63-221843 |
Sep 1988 |
JPX |
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Parent Case Info
This is a continuation of application Ser. No. 07/339,882, filed Aug. 29, 1989, now abandoned.
US Referenced Citations (3)
Number |
Name |
Date |
Kind |
4104125 |
Takechi et al. |
Aug 1978 |
|
4264738 |
Stepanov et al. |
Apr 1981 |
|
4374926 |
Stern |
Feb 1983 |
|
Foreign Referenced Citations (1)
Number |
Date |
Country |
0261941 |
Mar 1988 |
EPX |
Non-Patent Literature Citations (1)
Entry |
Scopes, "Protein Purification Principles and Practice", 2nd edition, 1987, pp. 110, 111, 122, 123. |
Continuations (1)
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Number |
Date |
Country |
Parent |
399882 |
Aug 1989 |
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