Patent US3475310A successfully refreshed from Google Patents.
US4096091A
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Limulus lysate having improved capacity to precipitate in the presence of low endotoxin concentrations, and reconstituting solutions therefor
Assignee: Baxter International Inc
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Filing: 1974-07-17
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Granted: 1978-06-20
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Expires: 1994-07-17
Abstract
Abstract A family of catalysts is disclosed for providing improved capacity for Limulus lysate solution to precipitate in the presence of extremely low concentrations of endotoxin. The catalyst family includes imidazole, manganese ions, Cleland's reagent, magnesium ions, strontium ions, barium ions, cysteine, lithium ions, and various combinations of the above materials. Solutions of the above catalysts may be used to advantage to reconstitute lyophilized Limulus lysate.
Inventors
- 👤 II Robert E. Hopkins
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-09 02:39
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/579
Claims Analysis
Total Claims: 56
Claim 1
Independent Claim
50 words
In a Limulus lysate solution, for providing improved capacity for the solution to precipitate in the presence of extremely low concentrations of endotoxin, the improvement comprising a catalytic concentration of a material selected from the group consisting of imidazole, ionic manganese, Cleland's reagent, glutathione, alkali metal thioglycollates, thiouracil, and cysteine.
Claim 2
Dependent Claim
50 words
In a Limulus lysate solution, for providing improved capacity for the solution to precipitate in the presence of extremely low concentrations of endotoxin, the improvement comprising a catalytic concentration of a material selected from the group consisting of imidazole, ionic manganese, Cleland's reagent, glutathione, alkali metal thioglycollates, thiouracil, and cysteine.
Claim 3
Dependent Claim
31 words
In a Limulus lysate solution, for providing improved capacity for the solution to precipitate in the presence of extremely low concentrations of endotoxin, the improvement comprising a catalytic concentration of imidazole.
Claim 4
Dependent Claim
31 words
In a Limulus lysate solution, for providing improved capacity for the solution to precipitate in the presence of extremely low concentrations of endotoxin, the improvement comprising a catalytic concentration of imidazole.
Claim 5
Dependent Claim
34 words
The Limulus lysate solution of claim 2 having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said lysate solution containing from 0.004 to 0.4 mole of imidazole per liter.
Claim 6
Dependent Claim
20 words
The solution of claim 3 in which from 0.01 to 0.3 mole of imidazole are present per liter of solution.
Claim 7
Dependent Claim
22 words
The solution of claim 3 in which from 0.005 to 0.3 mole of Li + ions are present per liter of solution.
Claim 8
Dependent Claim
22 words
The solution of claim 4 in which from 0.02 to 0.15 mole of Li + ions are present per liter of solution.
Claim 9
Dependent Claim
22 words
The solution of claim 3 in which from 0.005 to 0.3 mole of Mg ++ ions are present per liter of solution.
Claim 10
Dependent Claim
22 words
The solution of claim 4 in which from 0.01 to 0.1 mole of Mg ++ ions are present per liter of solution.
Claim 11
Dependent Claim
22 words
The solution of claim 3 in which from 0.0005 to 0.008 mole of [thioglycollate] - ions are present per liter of solution.
Claim 12
Dependent Claim
21 words
The solution of claim 9 in which said thioglycollate ions are added to said solution in the form of sodium thioglycollate.
Claim 13
Dependent Claim
22 words
The solution of claim 3 in which from 0.005 to 0.3 mole of Sr ++ ions are present per liter of solution.
Claim 14
Dependent Claim
22 words
The solution of claim 4 in which from 0.02 to 0.15 mole of Sr ++ ions are present per liter of solution.
Claim 15
Dependent Claim
22 words
The solution of claim 8 in which from 0.0005 to 0.008 mole of [thioglycollate] - ions are present per liter of solution.
Claim 16
Dependent Claim
37 words
The solution of claim 13 in which from 0.02 to 0.03 mole of Mg ++ ions; 0.02 to 0.03 mole of imidazole; and 0.001 to 0.002 mole of [thioglycollate] - ions are present per liter of solution.
Claim 17
Dependent Claim
19 words
The solution of claim 14 in which the Mg ++ ions are added in the form of magnesium chloride.
Claim 18
Dependent Claim
19 words
The solution of claim 15 in which said [thioglycollate] - ions are added in the form of sodium thioglycollate.
Claim 19
Dependent Claim
22 words
The solution of claim 3 in which from 0.01 to 0.2 mole of Li + ions are present per liter of solution.
Claim 20
Dependent Claim
19 words
The solution of claim 17 in which said Li + ions are added in the form of lithium chloride.
Claim 21
Dependent Claim
32 words
In a Limulus lysate solution, for providing improved capacity for the solution to precipitate in the presence of extremely low concentrations of endotoxin, the improvement comprising a catalytic concentration of ionic manganese.
Claim 22
Dependent Claim
32 words
In a Limulus lysate solution, for providing improved capacity for the solution to precipitate in the presence of extremely low concentrations of endotoxin, the improvement comprising a catalytic concentration of ionic manganese.
Claim 23
Dependent Claim
35 words
The Limulus lysate solution of claim 19 having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said lysate solution containing from 0.005 to 0.1 mole of ionic manganese per liter.
Claim 24
Dependent Claim
20 words
The solution of claim 20 in which from 0.01 to 0.04 mole of Mn ++ ions are present per liter.
Claim 25
Dependent Claim
19 words
The solution of claim 21 in which said Mn ++ ions are added in the form of manganese chloride.
Claim 26
Dependent Claim
32 words
In a Limulus lysate solution, for providing improved capacity for the solution to precipitate in the presence of extremely low concentrations of endotoxin, the improvement comprising a catalytic concentration of Cleland's reagent.
Claim 27
Dependent Claim
32 words
In a Limulus lysate solution, for providing improved capacity for the solution to precipitate in the presence of extremely low concentrations of endotoxin, the improvement comprising a catalytic concentration of Cleland's reagent.
Claim 28
Dependent Claim
37 words
The Limulus lysate solution of claim 23 having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said lysate solution containing from 0.00005 to 0.0005 mole of Cleland's reagent per liter of solution.
Claim 29
Dependent Claim
21 words
The solution of claim 24 in which from 0.0001 to 0.0003 mole of Cleland's reagent are present per liter of solution.
Claim 30
Dependent Claim
47 words
In a Limulus lysate solution, for providing improved capacity for the solution to precipitate in the presence of extremely low concentrations of endotoxin, the improvement comprising a catalytic concentration of an organic sulfhydryl-containing compound selected from the group consisting of cysteine, glutathione, alkali metal thioglycollates, and thiouracil.
Claim 31
Dependent Claim
47 words
In a Limulus lysate solution, for providing improved capacity for the solution to precipitate in the presence of extremely low concentrations of endotoxin, the improvement comprising a catalytic concentration of an organic sulfhydryl-containing compound selected from the group consisting of cysteine, glutathione, alkali metal thioglycollates, and thiouracil.
Claim 32
Dependent Claim
34 words
The Limulus lysate solution of claim 26 having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said lysate solution containing from 0.005 to 0.03 mole of cysteine per liter.
Claim 33
Dependent Claim
25 words
The solution of claim 27 in which said cysteine is added in the form of cysteine hydrochloride neutralized to generally neutral pH with an alkali.
Claim 34
Dependent Claim
14 words
The solution of claim 26 which also contains a catalytic amount of magnesium ions.
Claim 35
Dependent Claim
60 words
A Limulus lysate solution having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said lysate solution containing from 0.2 to 2 percent by weight of water-soluble magnesium salt and from 0.02 to 0.2 percent by weight of an organic sulfhydryl-containing compound selected from the group consisting of cysteine, alkali metal thioglycollate salts, and 2-thioamino uracil.
Claim 36
Dependent Claim
60 words
A Limulus lysate solution having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said lysate solution containing from 0.2 to 2 percent by weight of water-soluble magnesium salt and from 0.02 to 0.2 percent by weight of an organic sulfhydryl-containing compound selected from the group consisting of cysteine, alkali metal thioglycollate salts, and 2-thioamino uracil.
Claim 37
Dependent Claim
13 words
The solution of claim 30 in which said magnesium salt is magnesium chloride.
Claim 38
Dependent Claim
13 words
The solution of claim 31 in which said sulfhydryl-containing compound is sodium thioglycollate.
Claim 39
Dependent Claim
52 words
An endotoxin-free water solution for reconstituting lyophilized Limulus lysate to produce a solution having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said reconstituting solution containing from 0.004 to 0.4 mole of imidazole, and from 0.005 to 0.3 mole of Mg ++ ions per liter of solution.
Claim 40
Dependent Claim
52 words
An endotoxin-free water solution for reconstituting lyophilized Limulus lysate to produce a solution having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said reconstituting solution containing from 0.004 to 0.4 mole of imidazole, and from 0.005 to 0.3 mole of Mg ++ ions per liter of solution.
Claim 41
Dependent Claim
30 words
The reconstituting solution of claim 33 in which from 0.01 to 0.3 mole of imidazole and 0.01 to 0.1 mole of Mg ++ ions are present per liter of solution.
Claim 42
Dependent Claim
23 words
The reconstituting solution of claim 34 in which from 0.0001 to 0.008 mole of [thioglycollate] - ions are present per liter of solution.
Claim 43
Dependent Claim
22 words
The solution of claim 35 in which from 0.2 to 0.15 mole of Li + ions are present per liter of solution.
Claim 44
Dependent Claim
48 words
An endotoxin-free water solution for reconstituting lyophilized Limulus lysate to produce a solution having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said reconstituting solution containing from 0.004 to 0.4 mole of imidazole, and from 0.005 to 0.3 mole of Sr ++ ions.
Claim 45
Dependent Claim
48 words
An endotoxin-free water solution for reconstituting lyophilized Limulus lysate to produce a solution having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said reconstituting solution containing from 0.004 to 0.4 mole of imidazole, and from 0.005 to 0.3 mole of Sr ++ ions.
Claim 46
Dependent Claim
30 words
The solution of claim 37 in which from 0.01 to 0.3 mole of imidazole, and from 0.02 to 0.15 mole of Sr ++ ions are present per liter of solution.
Claim 47
Dependent Claim
69 words
An endotoxin-free water solution for reconstituting lyophilized Limulus lysate to provide a Limulus solution having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said reconstituting solution comprising from 0.2 to 2 percent by weight of water-soluble magnesium salt and from 0.02 to 0.2 percent by weight of a sulfhydryl-containing compound selected from the group consisting of cysteine, alkali metal thioglycollate salts, and 2-thio-amino uracil.
Claim 48
Dependent Claim
69 words
An endotoxin-free water solution for reconstituting lyophilized Limulus lysate to provide a Limulus solution having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said reconstituting solution comprising from 0.2 to 2 percent by weight of water-soluble magnesium salt and from 0.02 to 0.2 percent by weight of a sulfhydryl-containing compound selected from the group consisting of cysteine, alkali metal thioglycollate salts, and 2-thio-amino uracil.
Claim 49
Dependent Claim
13 words
The solution of claim 39 in which said magnesium salt is magnesium chloride.
Claim 50
Dependent Claim
27 words
The solution of claim 40 in which about 0.5 weight percent of magnesium chloride is present, and about 0.05 weight percent of said sulfhydryl-containing compound is present.
Claim 51
Dependent Claim
13 words
The solution of claim 41 in which said sulfhydryl-containing compound is sodium thioglycollate.
Claim 52
Dependent Claim
12 words
The solution of claim 41 in which said sulfhydryl-containing compound is cysteine.
Claim 53
Dependent Claim
50 words
An endotoxin-free water solution for reconstituting lyophilized Limulus lysate to provide a solution having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said reconstituting solution containing from 0.004 to 0.4 mole of imidazole and from 0.005 to 0.3 mole of Li + ions per liter.
Claim 54
Dependent Claim
50 words
An endotoxin-free water solution for reconstituting lyophilized Limulus lysate to provide a solution having improved capacity to precipitate in the presence of extremely low concentrations of endotoxin, said reconstituting solution containing from 0.004 to 0.4 mole of imidazole and from 0.005 to 0.3 mole of Li + ions per liter.
Claim 55
Dependent Claim
21 words
The solution of claim 44 in which from 0.03 to 0.15 mole of lithium chloride are present per liter of solution.
Claim 56
Dependent Claim
26 words
The solution of claim 32 in which about 0.5 percent by weight of magnesium chloride and about 0.05 percent by weight of sodium thioglycollate is present.
References Cited (Backward Citations)
US3954663A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1981-06-19 | 1981-12-19 | 1982-06-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1985-06-19 | 1985-12-19 | 1986-06-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1989-06-19 | 1989-12-19 | 1990-06-20 | $7700.00 | Window Closed / Lapsed |