US4468345A
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US
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Protein (PP17), a process for concentrating and isolating it and its use
Assignee: Siemens Healthcare Diagnostics GmbH Germany
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Filing: 1983-07-29
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Granted: 1984-08-28
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Expires: 2003-07-29
Abstract
Abstract The protein PP17, which has (a) an electrophoretic mobility in the range between beta 1- and alpha 2-globulins, (b) an isoelectric point of 5.25+/-0.20, (c) a sedimentation coefficient S20o, w of 2.7+/-0.1 S, (d) a molecular weight, determined in polyacrylamide gel containing sodium dodecyl sulfate (SDS), of 38,000+/-2,000, (e) an extinction coefficient E1cm1% (280 nm) of 8.5+/-0.4 and (f) a carbohydrate content of 2.1+/-0.9% containing 0.3+/-0.2% of mannose, 0.4+/-0.2% of galactose, 0.2+/-0.1% of xylose, 1.0+/-0.3% of N-acetyl-glucosamine and 0.2+/-0.1% of N-acetyl-neuramin acid, is described. A process for concentrating and isolating it and its use are also described.
Inventors
- 👤 Hans Bohn
- 👤 Wilhelm Winckler
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-27 14:07
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/46; C07K14/47; C07K14/4701; C07K14/4715; C07K16/00; C07K16/18; Y; Y10; Y10S; Y10S530/00; Y10S530/806; Y10S530/827; Y10S530/85; Y10S530/851
Claims Analysis
Total Claims: 6
Claim 1
Independent Claim
165 words
A protein, PP 17 , extracted from placental tissue, having (a) an electrophoretic mobility in the range between β 1 - and α 2 -globulins; (b) an isoelectric point of 5.25±0.20; (c) a sedimentation coefficient S 20 °.sub.,w of 2.7±0.1 S; (d) a molecular weight, determined in polyacrylamide gel containing sodium dodecyl sulfate (SDS), of 38,000±2,000; (e) an extinction coefficient E 1 cm 1% (280 nm) of 8.5±0.4; (f) a carbohydrate content of 2.1±0.9% containing 0.3±0.2% of mannose, 0.4±0.2% of galactose, 0.2±0.1% of xylose, 1.0±0.3% of N-acetyl-glycosamine and 0.2±0.1% of N-acetyl-neuramin acid; and (g) an amino acid analysis of ______________________________________
Variation
Amino Acid Mol % Coefficient
______________________________________
Lysine 5.86 5.52
Histidine 2.16 36.37
Arginine 5.30 2.90
Aspartic acid 7.44 2.59
Threonine 4.41 9.80
Serine 9.14 4.73
Glutamic acid 18.52 2.94
Proline 4.41 5.13
Glycine 4.93 2.92
Alanine 8.56 1.55
Cystine /2 0.75 3.58
Valine 6.91 3.53
Methionine 1.46 7.89
Isoleucine 2.53 4.70
Leucine 11.39 1.55
Tyrosine 1.43 11.97
Phenylalanine 2.79 7.29
Tryptophan 1.97 8.41.
______________________________________
Claim 2
Dependent Claim
165 words
A protein, PP 17 , extracted from placental tissue, having (a) an electrophoretic mobility in the range between β 1 - and α 2 -globulins; (b) an isoelectric point of 5.25±0.20; (c) a sedimentation coefficient S 20 °.sub.,w of 2.7±0.1 S; (d) a molecular weight, determined in polyacrylamide gel containing sodium dodecyl sulfate (SDS), of 38,000±2,000; (e) an extinction coefficient E 1 cm 1% (280 nm) of 8.5±0.4; (f) a carbohydrate content of 2.1±0.9% containing 0.3±0.2% of mannose, 0.4±0.2% of galactose, 0.2±0.1% of xylose, 1.0±0.3% of N-acetyl-glycosamine and 0.2±0.1% of N-acetyl-neuramin acid; and (g) an amino acid analysis of ______________________________________
Variation
Amino Acid Mol % Coefficient
______________________________________
Lysine 5.86 5.52
Histidine 2.16 36.37
Arginine 5.30 2.90
Aspartic acid 7.44 2.59
Threonine 4.41 9.80
Serine 9.14 4.73
Glutamic acid 18.52 2.94
Proline 4.41 5.13
Glycine 4.93 2.92
Alanine 8.56 1.55
Cystine /2 0.75 3.58
Valine 6.91 3.53
Methionine 1.46 7.89
Isoleucine 2.53 4.70
Leucine 11.39 1.55
Tyrosine 1.43 11.97
Phenylalanine 2.79 7.29
Tryptophan 1.97 8.41.
______________________________________
Claim 3
Dependent Claim
47 words
A process for isolating the protein PP 17 as claimed in claim 1 which comprises subjecting an extract from human placentas containing protein PP 17 to at least one known procedure for isolating proteins and, in each instance, recovering the material containing the protein to be isolated.
Claim 4
Dependent Claim
178 words
The process for concentrating the protein as claimed in claim 1, which comprises subjecting an extract from human placentas to at least one of the following measures and obtaining the PP 17 -enriched fraction: (1) precipitation with ammonium sulfate in the pH range from 5 to 8 and at 30-50% saturation; (2) precipitation with a water-soluble acridine base at a pH value between 7 and 9 and a concentration of the base of 0.4-0.8 g/100 ml; (3) precipitation of concomitant proteins in dilute salt solutions at pH 7.0 up to a concentration of 20% by volume of alcohol; (4) electrophoretic separation at pH 8.0 and isolation of the fraction with a mobility between that of β 1 - and α 2 -globulins; (5) isoelectric focusing in the pH range from 5.1 to 5.4; (6) dialysis against a buffer solution of pH 2 to 4 and removal of precipitated impurities; (7) gel filtration to obtain proteins with molecular weights from 20,000 to 60,000; (8) adsorption onto a weakly basic ion exchanger and elution of the protein; and (9) immunoadsorption.
Claim 5
Dependent Claim
17 words
The use of the protein as claimed in claim 1 for obtaining specific antibodies against this protein.
Claim 6
Dependent Claim
21 words
The use of the protein as claimed in claim 1 in immunological methods for the detection and determination of this protein.
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) | 1987-08-28 | 1988-02-27 | 1988-08-28 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-08-28 | 1992-02-27 | 1992-08-28 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-08-28 | 1996-02-27 | 1996-08-28 | $7700.00 | Window Closed / Lapsed |