US4431582A
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Protein, composition and method for enhancing epithelial cell movement
Assignee: Research Corp
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Filing: 1982-12-17
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Granted: 1984-02-14
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Expires: 2002-12-17
Abstract
Abstract A protein has been isolated from mammalian plasma or serum which enhances epithelial cell movement. A second factor which has no epithelial cell movement activity has also been isolated from mammalian plasma or serum which, when combined with the first protein, enhances the epithelial cell movement activity thereof.
Inventors
- 👤 Kurt S. Stenn
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 17:51
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/46; C07K14/47; C07K14/4701; C07K14/4702; C07K14/4705; C07K16/00; C07K16/18; A; A61; A61K; A61K38/00; Y; Y10; Y10S; Y10S530/00; Y10S530/827; Y10S530/829; Y10S530/83
Claims Analysis
Total Claims: 6
Claim 1
Independent Claim
199 words
A trypsin digestible single-chained, glycosylated protein which enhances epithelial cell movement in vivo having the following properties: (a) a molecular weight of about 62,000 daltons; (b) resistant to serine protease inhibitors, diisopropyl fluorophosphate and phenyl methyl sulfonyl fluoride; (c) isoelectric point between 5.1 and 5.2; (d) extinction coefficient of about 1.0 at 280 nm ultraviolet light, and (e) an amino acid composition including but not limited to: ______________________________________
No. of amino acids
No. of amino acid
Amino per protein molecule
molecules per 1000
Acid 62,752 daltons amino acids
______________________________________
Asp 62 111
Thr 40 72
Ser 46 81
Glu 67 119
Pro 19 35
Gly 26 46
Ala 43 77
Val 33 59
Met 15 26
Ile 24 44
Leu 69 123
Tyr 12 22
Phe 31 56
His 12 21
Lys 34 60
Arg 21 37
Trp 4 7
Cys 2 4;
______________________________________ (f) the N-terminal amino acid is asparagine and the N-terminal amino acid sequence is NH 2 -Asparagine-serine-proline-leucine-aspartic-glutamic; (g) immunoelectrophoresis properties of a serum α 2 globulin; (h) antibody to the protein blocks all the epidermal cell spreading activity of human serum; (i) antibody to the protein does not react with other known human serum proteins.
Claim 2
Dependent Claim
199 words
A trypsin digestible single-chained, glycosylated protein which enhances epithelial cell movement in vivo having the following properties: (a) a molecular weight of about 62,000 daltons; (b) resistant to serine protease inhibitors, diisopropyl fluorophosphate and phenyl methyl sulfonyl fluoride; (c) isoelectric point between 5.1 and 5.2; (d) extinction coefficient of about 1.0 at 280 nm ultraviolet light, and (e) an amino acid composition including but not limited to: ______________________________________
No. of amino acids
No. of amino acid
Amino per protein molecule
molecules per 1000
Acid 62,752 daltons amino acids
______________________________________
Asp 62 111
Thr 40 72
Ser 46 81
Glu 67 119
Pro 19 35
Gly 26 46
Ala 43 77
Val 33 59
Met 15 26
Ile 24 44
Leu 69 123
Tyr 12 22
Phe 31 56
His 12 21
Lys 34 60
Arg 21 37
Trp 4 7
Cys 2 4;
______________________________________ (f) the N-terminal amino acid is asparagine and the N-terminal amino acid sequence is NH 2 -Asparagine-serine-proline-leucine-aspartic-glutamic; (g) immunoelectrophoresis properties of a serum α 2 globulin; (h) antibody to the protein blocks all the epidermal cell spreading activity of human serum; (i) antibody to the protein does not react with other known human serum proteins.
Claim 3
Dependent Claim
177 words
A method of preparing the protein of claim 1 comprising: (1) diluting mammalian plasma or serum with water and dissolving therein sufficient ammonium sulfate to produce an approximately 33.3% saturated solution and removing the thus formed precipitate; (2) dissolving in the solution remaining from step (1) sufficient ammonium sulfate to produce an approximately 66.7% saturated solution; (3) removing the precipitate from the solution produced by step (2), suspending it in water and dialyzing the suspension against a buffered solution having a pH of about 6.0; (4) placing the dialyzed fraction in a buffered solution having a pH of about 6.0 and subjecting the solution to anionic chromatographic separation, including washing the chromatographically adhered fraction with said buffered solution; (5) eluting the chromatographically adhered fraction with a buffered solution having a pH of about 6.0 and an anionic strength of about 12 mmhos; (6) dialyzing the eluted fraction against a buffered solution having a pH of about 6.0; and (7) subjecting the dialysand to isoelectric precipitation at a pH of between 4.6 and 5.0 to produce the protein.
Claim 4
Dependent Claim
17 words
The method of claim 2 wherein said protein is further purified by gel filtration chromatography and electrophoresis.
Claim 5
Dependent Claim
127 words
A method of preparing a factor which enhances the epithelial cell movement activity of the protein of claim 2 comprising; (1) diluting mammalian plasma or serum with water and dissolving therein sufficient ammonium sulfate to produce an approximately 33.3% saturated solution and removing the thus formed precipitate; (2) dissolving in the solution remaining from step (1) sufficient ammonium sulfate to produce an approximately 66.7% saturated solution; (3) removing the precipitate from the solution produced by step (2), suspending it in water and dialyzing the suspension against a buffered solution having a pH of about 6.0; (4) placing the dialyzed fraction in a buffered solution having a pH of about 6.0 and subjecting the solution to anionic chromatographic separation, collecting the chromatographically non-adhered fraction with said buffered solution.
Claim 6
Dependent Claim
5 words
The product of claim 4.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-02-13 | 1987-08-15 | 1988-02-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-02-13 | 1991-08-15 | 1992-02-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-02-13 | 1995-08-15 | 1996-02-14 | $7700.00 | Window Closed / Lapsed |