US4624932A
Expired / Lapsed
US
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Recognins and their chemoreciprocals
Assignee: Individual
•
Filing: 1981-07-30
•
Granted: 1986-11-25
•
Expires: 2001-07-30
Abstract
Abstract A product, Astrocytin, derived from brain cancer cells growing in vivo, and the method of preparing same. Astrocytin may be complexed with an inert carrier, such as bromoacetyl cellulose, and employed in tests for brain tumors. The antibody to Astrocytin is also prepared. A complex of Astrocytin and an inert carrier may be further complexed to anti-Astrocytin.
Inventors
- 👤 Samuel Bogoch
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 07:56
CPC Classifications
C; C07; C07K; C07K14/00; C07K14/435; C07K14/46; C07K14/47; C07K14/4701; C07K14/4731; C07K16/00; C07K16/18; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/575; G01N33/5758; G01N33/57585; G01N2333/00; G01N2333/435; G01N2333/46; G01N2333/47; G01N2333/4701; G01N2333/473; Y; Y10; Y10S; Y10S530/00; Y10S530/81; Y10S530/812; Y10S530/813; Y10S530/814; Y10S530/827
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
198 words
A process for the production of a product, Astrocytin, which comprises extracting brain glioma tumor tissue with a neutral buffer by repeated disruption of the tissue to solubilize protein fractions, separating from the resulting extract of solubilized proteins the fraction having a pK range of from about 1 to 4, and isolating therefrom a product having a molecular weight of about 8000, said product, Astrocytin, characterized by forming a single line precipitate with its specific antibody in quantitative precipitin tests and Ouchterlony gel diffusion tests, being soluble in water and aqueous solutions having an acid or neutral pH, and insoluble at an alkaline pH, having a spectrophotometric absorption peak wave length of 280 mu, a molecular weight of about 8000 and further having an amino acid composition approximately as follows: ______________________________________
Approximate Number
of Residues
______________________________________
Aspartic Acid 9
Threonine 5
Serine 6
Glutamic Acid 13
Proline 4
Glycine 6
Alanine 9
Valine 4
1/2 Cysteine 2
Methionine 1
Isoleucine 2
Leucine 8
Tryosine 2
Phenylalanine 3
Lysine 8
Histidine 2
Arginine 4
Approximate Total = 88
______________________________________ the amino acids cysteic acid, hydroxyproline, norleucine, ammonia, isodesmosine, desmosine, hydroxylysine, lysinonorleucine and gamma-aminobutyric acid being absent in detectable amounts.
Claim 2
Dependent Claim
198 words
A process for the production of a product, Astrocytin, which comprises extracting brain glioma tumor tissue with a neutral buffer by repeated disruption of the tissue to solubilize protein fractions, separating from the resulting extract of solubilized proteins the fraction having a pK range of from about 1 to 4, and isolating therefrom a product having a molecular weight of about 8000, said product, Astrocytin, characterized by forming a single line precipitate with its specific antibody in quantitative precipitin tests and Ouchterlony gel diffusion tests, being soluble in water and aqueous solutions having an acid or neutral pH, and insoluble at an alkaline pH, having a spectrophotometric absorption peak wave length of 280 mu, a molecular weight of about 8000 and further having an amino acid composition approximately as follows: ______________________________________
Approximate Number
of Residues
______________________________________
Aspartic Acid 9
Threonine 5
Serine 6
Glutamic Acid 13
Proline 4
Glycine 6
Alanine 9
Valine 4
1/2 Cysteine 2
Methionine 1
Isoleucine 2
Leucine 8
Tryosine 2
Phenylalanine 3
Lysine 8
Histidine 2
Arginine 4
Approximate Total = 88
______________________________________ the amino acids cysteic acid, hydroxyproline, norleucine, ammonia, isodesmosine, desmosine, hydroxylysine, lysinonorleucine and gamma-aminobutyric acid being absent in detectable amounts.
Claim 3
Dependent Claim
42 words
The process of claim 1 wherein said step of separating the fraction having a pK range of from about 1 to 4 is carried out by adding said extract of solubilized proteins into a chromatographic column and eluting with increasingly acidic solvents.
Claim 4
Dependent Claim
30 words
The process of claim 2 wherein said isolating is carried out by filtering the eluate to obtain a fraction having Astrocytin and separating Astrocytin therefrom by thin layer gel chromatrography.
Claim 5
Dependent Claim
140 words
A product, Astrocytin, produced from brain tumor tissue, characterized by forming a single line precipitate with its specific antibody in quantitative precipitin tests and Ouchterlony gel diffusion tests, being soluble in water and aqueous solutions having an acid or neutral pH, and insoluble at an alkaline pH, having a spectrophotometric absorption peak wave length of 280 mu, a molecular weight of about 8000 and having an amino acid composition approximately as follows: ______________________________________
Approximate Number
of Residues
______________________________________
Aspartic Acid 9
Threonine 5
Serine 6
Glutamic Acid 13
Proline 4
Glycine 6
Alanine 9
Valine 4
1/2 Cysteine 2
Methionine 1
Isoleucine 2
Leucine 8
Tryosine 2
Phenylalanine 3
Lysine 8
Histidine 2
Arginine 4
Approximate Total = 88
______________________________________ the amino acids cysteic acid, hydroxyproline, norleucine, ammonia, isodesmosine, desmosine, hydroxylysine, lysinonorleucine and gamma-aminobutyric acid being absent in detectable amounts.
Claim 6
Dependent Claim
140 words
A product, Astrocytin, produced from brain tumor tissue, characterized by forming a single line precipitate with its specific antibody in quantitative precipitin tests and Ouchterlony gel diffusion tests, being soluble in water and aqueous solutions having an acid or neutral pH, and insoluble at an alkaline pH, having a spectrophotometric absorption peak wave length of 280 mu, a molecular weight of about 8000 and having an amino acid composition approximately as follows: ______________________________________
Approximate Number
of Residues
______________________________________
Aspartic Acid 9
Threonine 5
Serine 6
Glutamic Acid 13
Proline 4
Glycine 6
Alanine 9
Valine 4
1/2 Cysteine 2
Methionine 1
Isoleucine 2
Leucine 8
Tryosine 2
Phenylalanine 3
Lysine 8
Histidine 2
Arginine 4
Approximate Total = 88
______________________________________ the amino acids cysteic acid, hydroxyproline, norleucine, ammonia, isodesmosine, desmosine, hydroxylysine, lysinonorleucine and gamma-aminobutyric acid being absent in detectable amounts.
Claim 7
Dependent Claim
13 words
The antibody, Anti-Astrocytin, produced in response to the product, Astrocytin, of claim 4.
Claim 8
Dependent Claim
11 words
The product of claim 4 complexed with an inert carrier substance.
Claim 9
Dependent Claim
12 words
The product of claim 6 wherein said inert carrier substance is bromoacetylcellulose.
Claim 10
Dependent Claim
11 words
The product of claim 5 complexed with an inert carrier substance.
Claim 11
Dependent Claim
12 words
The product of claim 8 wherein said inert carrier substance is bromoacetylcellulose.
Claim 12
Dependent Claim
12 words
The product of claim 6 further complexed with the antibody to Astrocytin.
Claim 13
Dependent Claim
12 words
The product of claim 7 further complexed with the antibody to Astrocytin.
Claim 14
Dependent Claim
141 words
A process for producing a product, Slow-Target Attaching Globulin (S-TAG), which comprises reacting blood serum or other body fluid with the product of claim 4 complexed with an inert carrier substance, for approximately two hours or more at a low temperature of about 4° C. to prevent degradative reactions and treating the resulting material with dilute acid to cleave S-TAG therefrom, said product S-TAG characterized by being soluble in aqueous buffered solutions, forming a single line precipitate with the product of claim 4 in Ouchterlony gel diffusion tests, being non-dialyzable in cellophane membranes, being retained by millipore filters which retain molecules over 25,000 molecular weight, having molecular weights in different stages of aggregations as determined by thin layer gel chromatography of approximately 50,000, and multiples thereof into the macroglobulin range and having a spectrophotometric absorption peak wave length of 280 mu.
Claim 15
Dependent Claim
12 words
The process of claim 12 wherein said inert carrier substance is bromoacetylcellulose.
Claim 16
Dependent Claim
138 words
A process for producing a product, Fast-Target Attaching Globulin (F-TAG), which comprises reacting blood serum or other body fluid with the product of claim 4 complexed with an inert carrier substance, for approximately 10 minutes at low temperature to of about 4° C. prevent degradative reactions and treating the resulting material with dilute acid to cleave F-TAG therefrom, said product F-TAG characterized by being soluble in aqueous buffered solutions, forming a single line precipitate with the product of claim 4 in Ouchterlony gel diffusion tests, being non-dialyzable in cellophane membranes, being retained by millipore filters which retain molecules over 25,000 molecular weight, having molecular weights in different stages of aggregation as determined by thin layer gel chromatography of approximately 50,000, and multiples thereof into the macroglobulin range and having a spectrophotometric absorption peak wave length of 280 mu.
Claim 17
Dependent Claim
12 words
The process of claim 14 wherein said inert carrier substance is bromoacetylcellulose.
Claim 18
Dependent Claim
43 words
A polypeptide product having the physicochemical specificities of the product of claim 4 (Astrocytin) and having a molecular weight approximately defined by the formula 4000+200X=Y wherein Y is te molecular weight of the product and X is an integer from 0 to 19.
Claim 19
Independent Claim
322 words
A method of detecting malignant cancer tumors in living mammals which comprises: a. dividing a sample of body fluid into a first and second aliquot; b. contacting said first aliquot with a complex of Astrocytin and an inert carrier for about two hours to allow protein to bind to said complex; c. separating from said first aliquot said complex and the protein bound to it; d. cleaving the bound protein from said complex; e. quantitatively determining the amount of protein cleaved from said complex; f. contacting said second aliquot with a complex of Astrocytin and an inert carrier for about 10 minutes to allow protein to bind to said complex; g. separating from said second aliquot said complex and the protein bound to it; h. cleaving the bound protein from said complex; i. quantitatively determining the amount of protein cleaved from said complex; j. comparing the amount of protein determined in step e, above, with that determined in step i, above; and correlating the difference between the two amounts with a predetermined standard value indicative of the tumor, wherein said Astrocytin is a product derived from brain tumor cells and is characterized by forming a single line precipitate with its specific antibody in quantitative precipitin tests and in Ouchterlony gel diffusion tests, being soluble in water and aqueous solutions having an acid or neutral pH, and insoluble at an alkaline pH, having a spectrophotometric absorption peak wave length of 280 mu, a molecular weight of about 8,000 and having an amino acid composition approximately as follows: ______________________________________
Approx. No.
of Residues
______________________________________
Aspartic Acid 9
Threonine 5
Serine 6
Glutamic Acid 13
Proline 4
Glycine 6
Alanine 9
Valine 4
1/2 Cystine 2
Methionine 1
Isoleucine 2
Leucine 8
Tryosine 2
Phenylalanine 3
Lysine 8
Histidine 2
Arginine 4
Approximate Total = 88
______________________________________ the amino acids cysteic, hydroxyproline, norleucine, ammonia, isodesmosine, hydroxylysine, lysinonorleucine and gamma-aminobutyric acid being absent in detectable amounts.
Claim 20
Independent Claim
322 words
A method of detecting malignant cancer tumors in living mammals which comprises: a. dividing a sample of body fluid into a first and second aliquot; b. contacting said first aliquot with a complex of Astrocytin and an inert carrier for about two hours to allow protein to bind to said complex; c. separating from said first aliquot said complex and the protein bound to it; d. cleaving the bound protein from said complex; e. quantitatively determining the amount of protein cleaved from said complex; f. contacting said second aliquot with a complex of Astrocytin and an inert carrier for about 10 minutes to allow protein to bind to said complex; g. separating from said second aliquot said complex and the protein bound to it; h. cleaving the bound protein from said complex; i. quantitatively determining the amount of protein cleaved from said complex; j. comparing the amount of protein determined in step e, above, with that determined in step i, above; and correlating the difference between the two amounts with a predetermined standard value indicative of the tumor, wherein said Astrocytin is a product derived from brain tumor cells and is characterized by forming a single line precipitate with its specific antibody in quantitative precipitin tests and in Ouchterlony gel diffusion tests, being soluble in water and aqueous solutions having an acid or neutral pH, and insoluble at an alkaline pH, having a spectrophotometric absorption peak wave length of 280 mu, a molecular weight of about 8,000 and having an amino acid composition approximately as follows: ______________________________________
Approx. No.
of Residues
______________________________________
Aspartic Acid 9
Threonine 5
Serine 6
Glutamic Acid 13
Proline 4
Glycine 6
Alanine 9
Valine 4
1/2 Cystine 2
Methionine 1
Isoleucine 2
Leucine 8
Tryosine 2
Phenylalanine 3
Lysine 8
Histidine 2
Arginine 4
Approximate Total = 88
______________________________________ the amino acids cysteic, hydroxyproline, norleucine, ammonia, isodesmosine, hydroxylysine, lysinonorleucine and gamma-aminobutyric acid being absent in detectable amounts.
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) | 1989-11-24 | 1990-05-26 | 1990-11-25 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1993-11-24 | 1994-05-26 | 1994-11-25 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1997-11-24 | 1998-05-26 | 1998-11-25 | $7700.00 | Window Closed / Lapsed |