US3948875A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Process for the preparation of epidermal growth factor and new derivative using cross-linked polyacrylamide gel at a pH of 1-3
Assignee: Individual
•
Filing: 1973-11-27
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Granted: 1976-04-06
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Expires: 1993-11-27
Abstract
Abstract A new procedure has been devised for the preparation of pure epidermal growth factor (EGF) in high yield which involves passing homogenates of submaxillary glands of adult male mice through a polyacrylamide gel column of a low acid pH. Preferably a two step procedure is used wherein the fraction obtained from the polyacrylamide gel chromatography is subjected to DEAE-cellulose chromatography. The new procedure also provides a new derivative of EGF, lacking the COOH-terminal Leu-Arg residues.
Inventors
- 👤 Stanley Cohen
- 👤 C. Richard Savage, Jr.
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-23 18:29
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/485; Y; Y10; Y10S; Y10S530/00; Y10S530/827; Y10S530/854
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
83 words
A process for the preparation of substantially pure epidermal growth factors which comprises homogenizing submaxillary glands of adult male mice, separating the resulting homogenate into a solid fraction and a supernatant fraction, passing the supernatant fraction through a column of crosslinked polyacrylamide gel at a pH of 1 to 3 to selectively adsorb epidermal growth factor from said supernatant fraction, eluting the column to recover a fractional eluate having epidermal growth activity and absorption of ultraviolet light of wave length about 280 μ.
Claim 2
Dependent Claim
83 words
A process for the preparation of substantially pure epidermal growth factors which comprises homogenizing submaxillary glands of adult male mice, separating the resulting homogenate into a solid fraction and a supernatant fraction, passing the supernatant fraction through a column of crosslinked polyacrylamide gel at a pH of 1 to 3 to selectively adsorb epidermal growth factor from said supernatant fraction, eluting the column to recover a fractional eluate having epidermal growth activity and absorption of ultraviolet light of wave length about 280 μ.
Claim 3
Dependent Claim
11 words
The process of claim 1 wherein said pH is about 1.5.
Claim 4
Dependent Claim
16 words
The process of claim 2 wherein the elution is effected with a hydrogen chloride-sodium chloride buffer.
Claim 5
Dependent Claim
18 words
The process of claim 1 wherein the pH of the homogenate is maintained at between 3 and 4.
Claim 6
Dependent Claim
32 words
The process of claim 1 wherein the pH of the homogenate is above 4 during said separation and the supernatant fraction is adjusted to a pH of between about 1 and 3.
Claim 7
Dependent Claim
20 words
The process of claim 1 wherein the crosslinked polyacrylamide gel has an exclusion limit of about 10,000 to 30,000 daltons.
Claim 8
Dependent Claim
15 words
The process of claim 5 wherein the pH of the supernatant fraction is about 1.5.
Claim 9
Dependent Claim
17 words
The process of claim 7 wherein the crosslinked polyacrylamide gel has an exclusion limit of 20,000 daltons.
Claim 10
Dependent Claim
148 words
A process for the production of high yields of pure epidermal growth factors which comprises homogenizing submaxillary glands of adult male mice, separating the resulting homogenate into a solid fraction and a supernatant fraction, passing the supernatant fraction through a column of crosslinked polyacrylamide gel at a pH of about 1 to 3 to selectively adsorb epidermal growth factors from said supernatant fraction, eluting the column to recover a fractional eluate having epidermal growth activity and adsorption of ultraviolet light of wave length about 280 mu, passing said fractional eluate through a column of diethylaminoethylcellulose at a pH of above about 4 to adsorb said epidermal growth factors therefrom and eluting the diethylaminoethylcellulose column with an increasing salt gradient at a pH of above about 4 to recover fractional eluates of increased purity having epidermal growth activity and absorption of ultraviolet light of wave length about 280 μ.
Claim 11
Dependent Claim
148 words
A process for the production of high yields of pure epidermal growth factors which comprises homogenizing submaxillary glands of adult male mice, separating the resulting homogenate into a solid fraction and a supernatant fraction, passing the supernatant fraction through a column of crosslinked polyacrylamide gel at a pH of about 1 to 3 to selectively adsorb epidermal growth factors from said supernatant fraction, eluting the column to recover a fractional eluate having epidermal growth activity and adsorption of ultraviolet light of wave length about 280 mu, passing said fractional eluate through a column of diethylaminoethylcellulose at a pH of above about 4 to adsorb said epidermal growth factors therefrom and eluting the diethylaminoethylcellulose column with an increasing salt gradient at a pH of above about 4 to recover fractional eluates of increased purity having epidermal growth activity and absorption of ultraviolet light of wave length about 280 μ.
Claim 12
Dependent Claim
24 words
The process of claim 9 wherein the supernatant fraction is passed through the column of crosslinked polyacrylamide gel at a pH of about 1.5
Claim 13
Dependent Claim
17 words
The process of claim 10 wherein the pH of the homogenate is between about 1 and 3.
Claim 14
Dependent Claim
16 words
The process of claim 9 wherein the pH of the homogenate is between 3 and 4.
Claim 15
Dependent Claim
31 words
The process of claim 9 wherein the pH of the homogenate is above 4 during said separation and the supernatant fraction is adjusted to a pH between about 1 and 3.
Claim 16
Dependent Claim
19 words
The process of claim 9 wherein the crosslinked polyacrylamide gel has an exclusion limit of 10 to 30,000 daltons.
Claim 17
Dependent Claim
32 words
The process of claim 14 wherein the homogenate is maintained at a pH of 1 to 3 or above 4 and the crosslinked polyacrylamide gel has an exclusion limit of 20,000 daltons.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1979-04-06 | 1979-10-06 | 1980-04-06 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1983-04-06 | 1983-10-06 | 1984-04-06 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1987-04-06 | 1987-10-06 | 1988-04-06 | $7700.00 | Window Closed / Lapsed |