Claims
- 1. A process for producing substantially pure chondroitinase ABC enzyme, said process comprising the steps of:
- (a) suspending cells of a Proteus vulgaris ATCC 6896 in a buffer solution and lysing the cells to obtain a crude enzyme-containing extract
- (b) removing nucleic acids from the enzyme-containing extract,
- (c) contacting the enzyme-containing extract obtained in step (b) with a first cation exchange resin, thereby adsorbing the enzyme,
- (d) eluting the adsorbed enzyme,
- (e) contacting the eluate with a second cation exchange resin, thereby adsorbing the enzyme, and
- (f) eluting the adsorbed enzyme; wherein the enzyme obtained after step (f) is substantially free of protease activity and nucleic acids.
- 2. The process of claim 1 wherein the first cation exchange resin contains sulfoalkyl exchange groups.
- 3. The process of claim 1 wherein the second cation exchange resin contains sulfoalkyl exchange groups.
- 4. The process of claim 1 wherein the buffer solution in step (a) further comprises a surfactant.
- 5. A process for preparing substantially pure chondroitinase ABC comprising,
- (a) contacting cells of Proteus vulgaris ATCC 6896 with a surfactant solution under conditions sufficient to lyse the cells, thereby producing an enzyme-containing extract,
- (b) removing nucleic acids from the enzyme-containing extract;
- (c) contacting the enzyme-containing extract with a cation exchange resin to adsorb the chondroitinase ABC to said resin, and
- (d) eluting the chondroitinase ABC adsorbed to said resin while continuously changing the ionic strength of the eluent; wherein the chondroitinase ABC produced after step (d) is substantially free of protease activity and nucleic acids.
- 6. The process of claim 5 wherein the cation exchange resin contains carboxyalkyl exchange groups.
- 7. The process of claim 5 wherein the surfactant is a nonionic surfactant.
Priority Claims (2)
Number |
Date |
Country |
Kind |
4-192882 |
Jun 1992 |
JPX |
|
4-310980 |
Oct 1992 |
JPX |
|
Parent Case Info
This is a divisional of application Ser. No. 08/082,853, filed Jun. 23, 1993, now U.S. Pat. No. 5,496,718.
US Referenced Citations (5)
Foreign Referenced Citations (1)
Number |
Date |
Country |
3080079 |
Aug 1987 |
JPX |
Divisions (1)
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Number |
Date |
Country |
Parent |
82853 |
Jun 1993 |
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