US4444760A
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Purification and characterization of a protein fibroblast growth factor
Assignee: Merck and Co Inc
•
Filing: 1983-06-17
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Granted: 1984-04-24
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Expires: 2003-06-17
Abstract
Abstract Acidic brain fibroblast growth factor isolated from bovine brain, is purified by a particular combination of protein purification techniques. The product is useful in the promotion of cell division (mitogenesis) such as in the promotion of wound healing.
Inventors
- 👤 Kenneth A. Thomas, Jr.
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 20:45
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/50; C07K14/501; A; A61; A61P; A61P43/00; A61K; A61K38/00; Y; Y10; Y10S; Y10S530/00; Y10S530/827; Y10S530/839
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
70 words
An acidic brain fibroblast growth factor in substantially pure form with molecular mass of 16,600 or 16,800 daltons and an amino acid composition of: ______________________________________
Amino Acid Units
______________________________________
Aspartic Acid
14
Asparagine
Threonine 9
Serine 10
Glutamic Acid
16
Glutamine
Proline 7
Glycine 14
Alanine 5
Cysteine 4
Valine 5
Methionine 1
Isoleucine 6
Leucine 19
Tyrosine 7
Phenylalanine 7
Histidine 5
Lysine 13
Arginine 6
Tryptophan 1
______________________________________
Claim 2
Dependent Claim
70 words
An acidic brain fibroblast growth factor in substantially pure form with molecular mass of 16,600 or 16,800 daltons and an amino acid composition of: ______________________________________
Amino Acid Units
______________________________________
Aspartic Acid
14
Asparagine
Threonine 9
Serine 10
Glutamic Acid
16
Glutamine
Proline 7
Glycine 14
Alanine 5
Cysteine 4
Valine 5
Methionine 1
Isoleucine 6
Leucine 19
Tyrosine 7
Phenylalanine 7
Histidine 5
Lysine 13
Arginine 6
Tryptophan 1
______________________________________
Claim 3
Dependent Claim
13 words
The acidic brain fibroblast growth factor of claim 1, isolated from bovine brain.
Claim 4
Dependent Claim
49 words
A process for the isolation of acidic brain fibroblast growth factor in substantially pure form which comprises the sequence of steps of: (1) extraction and differential salt precipitation from the source tissue; (2) ion exchange; (3) gel filtration; (4) ion exchange; (5) isoelectric focusing; (6) hydrophobic reverse phase chromatography.
Claim 5
Dependent Claim
49 words
A process for the isolation of acidic brain fibroblast growth factor in substantially pure form which comprises the sequence of steps of: (1) extraction and differential salt precipitation from the source tissue; (2) ion exchange; (3) gel filtration; (4) ion exchange; (5) isoelectric focusing; (6) hydrophobic reverse phase chromatography.
Claim 6
Dependent Claim
11 words
The process of claim 3 for the isolation from bovine brain.
Claim 7
Dependent Claim
196 words
The process of claim 3 wherein: (1) The extraction and differential salt precipitation is accomplished by sequential treatments with 0.15 M ammonium sulfate at pH 4.5; 1.52 M ammonium sulfate at pH 6.75; and 3.41 M ammonium sulfate at pH 6.75 followed by dialysis and lyophilization of the high salt precipitate; (2) the ion exchange comprises batch adsorption on carboxymethyl-Sephadex (Pharmacia) resin at pH 6 followed by sequential elution with 0.1 M sodium phosphate buffer containing 0.15 M and 0.60 M sodium chloride followed by dialysis and lyophilization of the 0.60 M sodium chloride fraction, (3) gel filtration on a Sephadex G-75 (Pharmacia) column by elution with 0.1 M ammonium bicarbonate, pH 8.5 followed by lyophilization of the pooled high activity fractions; (4) ion exchange on a carboxymethyl cellulose column loaded in 0.1 M ammonium formate, pH 6.0 and eluted with 0.2 M and 0.6 M ammonium formate, pH 6.0, in sequence followed by lyophilization of the active pool; (5) isoelectric focusing is in Ultrodex (LKB, Gaithersburg, Md.); and (6) hydrophobic reverse phase high performance liquid chromatographic purification of the acidic mitogen is on Vydac C 4 silica based HPLC column (The Separations Group, Hesperia, Calif.).
Claim 8
Dependent Claim
11 words
The process of claim 5 for the isolation from bovine brain.
Claim 9
Dependent Claim
23 words
A pharmaceutical growth promoting composition comprising a pharmaceutical carrier and an effective growth promoting amount of the fibroblast growth factor of claim 1.
Claim 10
Dependent Claim
32 words
A method of promoting wound healing which comprises the administration to a patient in need of such treatment of an effective growth promoting amount of the fibroblast growth factor of claim 1.
References Cited (Backward Citations)
US4195017A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-04-24 | 1987-10-24 | 1988-04-24 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-04-24 | 1991-10-24 | 1992-04-24 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-04-24 | 1995-10-24 | 1996-04-24 | $7700.00 | Window Closed / Lapsed |