Claims
- 1. Process for production of a thrombolytically active protein which consists of amino acid sequence:
- ______________________________________ 1 SYQGNSDCYF GNGSAYRGTH SLTHSGASCL PWNSMILIGK VYTAQNPSAQ 51 ALGLGKHNYC RNPDGDAKPW CHVLKNRRLT WEYCDVPSCS TQGLRQYSQP101 QFRIKGGLFA DIASHPWQAA IFAKHRRSPG ERFLCGGILI SSCWILSAAH151 CFQERFPPHH LTVILGRTYR VVPGEEEQKF EVEKYIVHKE FDDDTYDNDI201 ALLQLKSDSS RCAQESSVVR TVCLPPADLQ LPDWTECELS GYGKHEALSP251 FYSERLKFAH VRLYPSSRCT SQHLLNRYVT DNMLCAGDTR SGGPQANLHD301 ACQGDSGGPL VCLNDGRMTL VGHSWGLGC GQKDVPGVYT XVTNYLDWIR351 DNMRP______________________________________
- comprising transforming a host cell with a plasmid which comprises a nucleic acid molecule which encodes said thrombolytically active protein, culturing said host cell to produce said thrombolytically active protein, and isolating said thrombolytically active protein following its production.
- 2. The process of claim 1, further comprising lysing said host cell prior to isolating said thrombolytically active protein.
- 3. The process of claim 1, wherein said host cell is a prokaryotic host cell.
- 4. The process of claim 3, wherein said prokaryotic host cell is an E. coli host cell.
- 5. Process for producing a thrombolytically active protein which consists of amino acid sequence:
- ______________________________________ 1 SYQGNSDCYF GNGSAYRGTH SLTESGASCL PWNSMILIGK VYTAQNPSAQ 51 ALGLGKHNYC RNPDGDAKPW CHVLKNRRLT WEYCDVPSCS TCGLRQYSQP101 QFRIKGGLFA DIASHPWQAA IFAKHRRSPG ERFLCGGILI SSCWILSAAH151 CFQERFPPHH LTVILGRTYR VVPGEEEQKF EVEKYIVHKE FDDDTYDNDI201 ALLQLKSDSS RCAQESSVVR TVCLPPADLQ LPDWTECELS GYGKHEALSP251 FYSERLKEAH VRLYPSSRCT SQHLLNRTVT DNMLCAGDTR SGGPQANLHD301 ACQGDSGGPL VCLNDGRMTL VGHSWGLGC GQKDVPGVTT KVTNYLDWIR351 DNMRP______________________________________
- comprising culturing a host cell which produces said thrombolytically active protein in the form of inclusion bodies to produce said inclusion bodies, separating said inclusion bodies from said host cell, solubilizing said inclusion bodies with guanidine hydrochloride, contacting solubilized inclusion bodies with oxidized glutathione, and renaturing protein with L-arginine and GSH.
- 6. The process of claim 5, further comprising contacting said thrombolytically active protein to an affinity chromatography column.
- 7. The process of claim 6, wherein said affinity chromatography column is an Erythrina-Trypsin-Inhibitor adsorber column.
- 8. The process of claim 6, further comprising concentrating said thrombolytically active protein prior to contact to said affinity chromatography column.
- 9. The process of claim 6, further comprising eluting said thrombolytically active protein from said affinity chromatography column.
- 10. The process of claim 6, comprising eluting said thrombolytically active protein with a solution having a pH of from 3 to 5.5.
Priority Claims (1)
Number |
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39 03 581.6 |
Feb 1989 |
DEX |
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RELATED APPLICATIONS
This application is a divisional of U.S. Ser. No. 08/217,617 filed Mar. 25, 1994, now U.S. Pat. No. 5,676,947, which is a continuation-in-part of two U.S. applications, one of which is U.S. Ser. No. 08/165,577 filed Dec. 13, 1993, abandoned, which is a continuation of U.S. Ser. No. 07/892,629 filed Jun. 2, 1992, abandoned, which is a continuation of U.S. Ser. No. 07/527,498 filed May 23, 1990, abandoned; and the other of which is U.S. Ser. No. 08/130,005 filed Sep. 30, 1993, abandoned, which is a divisional of U.S. Ser. No. 07/968,171 filed Oct. 29, 1992, abandoned, which is a continuation of U.S. Ser. No. 07/585,129, filed as PCT/EP90/00194, Feb. 6, 1990, now U.S. Pat. No. 5,223,256.
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Divisions (2)
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217617 |
Mar 1994 |
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968171 |
Oct 1992 |
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Continuations (3)
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892629 |
Jun 1992 |
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527498 |
May 1990 |
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585129 |
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Continuation in Parts (2)
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130005 |
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