Claims
- 1. A method for the isolation of an essentially purified and isolated heparin-binding brain mitogen having the N-terminal amino acid sequence H-Gly-Lys-Lys-Glu-Lys-Pro-Glu-Lys-Lys-Val-, wherein said mitogen is selected from the group consisting of HBBM-1, HBBM-2 and HBBM-3, wherein HBBM-1 has a molecular weight of about 18,000 daltons, HBBM-2 has a molecular weight of about 16,000 daltons and HBBM-3 has a molecular weight of about 15,000 daltons, where said molecular weights are determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis under either reducing or non-reducing conditions, which comprises the sequence of steps of:
- (a) extraction from the source tissue to obtain an extract containing the heparin-binding brain mitogen;
- (b) subjecting the extract from step (a) to a first cation-exchange chromatography, such that the eluate contains the heparin-binding brain mitogen;
- (c) subjecting the eluate from step (b) to heparin affinity chromatography, wherein the eluate is added to a heparin affinity column, such that the heparin-binding brain mitogen binds to the column and is then eluted from the column; and
- (d) subjecting the eluate from step (c) to a second cation-exchange chromatography to separate any contaminating acidic fibroblast growth factor from the heparin-binding brain mitogen.
- 2. The method of claim 1 which further comprises the step of hydrophobic-interaction chromatography after step (d).
- 3. The method of claim 1 wherein the eluted fractions of steps (c) and (d) are analyzed for the presence of heparin-binding brain mitogen by reverse-phase high performance liquid chromatography.
- 4. The method of claim 3 wherein the source tissue is bovine brain.
Parent Case Info
This is a divisional of co-pending application Ser. No. 07/787,692, filed on Nov. 1, 1991, U.S. Pat. No. 5,171,842, which is a continuation of application Ser. No. 07/147,835, filed on Jan. 25, 1988, now abandoned.
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Divisions (1)
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Date |
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Parent |
787692 |
Nov 1991 |
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Continuations (1)
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197835 |
Jan 1988 |
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