US3997404A
Expired / Lapsed
US
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Method and apparatus for characterizing biologically active agents
Assignee: Johnston Laboratories Inc
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Filing: 1975-05-27
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Granted: 1976-12-14
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Expires: 1995-05-27
Abstract
Abstract A method for characterization of biological activity comprising providing a plurality of gas-tight cells each having a light-permeable wall section, each cell containing a different 14 C-labeled fermentable substrate, a carbon dioxide absorber and a scintillation compound in intimate proximity to each other remote from said substrate, and an axenic sample of a biologically active material; subjecting said cells to conditions conducive to biological activity and thereafter measuring the light emitted by the scintillation compound as a result of the decay of 14 CO 2 generated by the metabolic action of the biologically active agent in said sample on said substrate and captured by the carbon dioxide absorber. Apparatus is provided comprising a plurality of sterile chambers containing different 14 C-labeled fermentable substrates, means to facilitate introduction of an axenic sample of a biologically active agent into each chamber, means for sealing each chamber to form a gas-tight cell having a light-permeable wall section, and a carbon dioxide absorber and a scintillation compound in intimate proximity to each other in each cell adjacent the light-permeable wall section.
Inventors
- 👤 John R. Waters
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 15:56
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/04; C12Q1/16; C12M; C12M41/00; C12M41/30; C12M41/34; C12M41/36; G; G01; G01N; G01N21/00; G01N21/75; G01N21/76; Y; Y10; Y10S; Y10S435/00; Y10S435/808
Claims Analysis
Total Claims: 51
Claim 1
Independent Claim
111 words
Apparatus for characterization of a biologically active agent comprising: a. a plurality of sterile chambers containing different 14 C-labeled fermentable substrates, b. means to facilitate introduction of an axenic sample of a biologically active agent to be characterized into each chamber, c. means for sealing each chamber to form a gas-tight cell having a light permeable wall section, d. a carbon dioxide absorber in each cell, and e. a scintillation compound in each cell in intimate proximity to said carbon dioxide absorber; said carbon dioxide absorber and said scintillation compound in each cell being disposed adjacent said light permeable wall section and out of contact with said 14 C-labeled fermentable substrates.
Claim 2
Dependent Claim
111 words
Apparatus for characterization of a biologically active agent comprising: a. a plurality of sterile chambers containing different 14 C-labeled fermentable substrates, b. means to facilitate introduction of an axenic sample of a biologically active agent to be characterized into each chamber, c. means for sealing each chamber to form a gas-tight cell having a light permeable wall section, d. a carbon dioxide absorber in each cell, and e. a scintillation compound in each cell in intimate proximity to said carbon dioxide absorber; said carbon dioxide absorber and said scintillation compound in each cell being disposed adjacent said light permeable wall section and out of contact with said 14 C-labeled fermentable substrates.
Claim 3
Dependent Claim
23 words
Apparatus as recited in claim 1 further comprising means to measure light emitted by said scintillation compound through said light permeable wall section.
Claim 4
Dependent Claim
21 words
Apparatus as recited in claim 2 further comprising light guide means communicating between said transparent wall section and said measuring means.
Claim 5
Dependent Claim
13 words
Apparatus as recited in claim 2 wherein said measuring means comprises a photomultiplier.
Claim 6
Dependent Claim
25 words
Apparatus as recited in claim 2 further comprising means to move said measuring means successively into alignment with the transparent wall section of each cell.
Claim 7
Dependent Claim
17 words
Apparatus as recited in claim 2 further comprising means to record values measured by said measuring means.
Claim 8
Dependent Claim
41 words
A device as recited in claim 1 wherein said 14 C-labeled fermentable substrates are selected from the class consisting of monosaccharides, disaccharides, polysaccharides, amino acids, salts of carboxylic acids, dicarboxylic acids and salts thereof, glycerol, polyhydroxy alcohols, hydroxy-carboxylic acids, and uracil.
Claim 9
Dependent Claim
11 words
Apparatus as recited in claim 1 wherein said chambers are sterilizable.
Claim 10
Dependent Claim
16 words
Apparatus as recited in claim 1 wherein said chambers are formed in a single integral body.
Claim 11
Dependent Claim
22 words
Apparatus as recited in claim 1 wherein said cavities are formed of opaque material and said sealing means comprises a transparent cover.
Claim 12
Dependent Claim
32 words
Apparatus as recited in claim 10 wherein said cover comprises a flat sheet of transparent material, and a gasket means interposed between said transparent sheet and said 14 C-labeled substrate containing chambers.
Claim 13
Dependent Claim
12 words
Apparatus as recited in claim 11 wherein said transparent cover is glass.
Claim 14
Dependent Claim
32 words
Apparatus as recited in claim 11 wherein each chamber has an opening and said sheet of transparent material is provided with a recess aligned to register with the opening of each chamber.
Claim 15
Dependent Claim
23 words
Apparatus as recited in claim 13 wherein said cover is provided with optical absorption capacity between cells in order to prevent cross talk.
Claim 16
Dependent Claim
20 words
Apparatus as recited in claim 13 wherein said recesses are coated with a scintillation compound and a carbon dioxide absorber.
Claim 17
Dependent Claim
14 words
Apparatus as recited in claim 15 wherein said carbon dioxide absorber is sodium hydroxide.
Claim 18
Dependent Claim
20 words
Apparatus as recited in claim 1 wherein said carbon dioxide absorber comprises an alkali metal or alkaline earth metal hydroxide.
Claim 19
Dependent Claim
18 words
Apparatus as recited in claim 1 wherein said scintillation compound comprises a mixture of 2,5-diphenyloxazole and p-bis(2-(5-phenyloxazoyl)) benzene.
Claim 20
Dependent Claim
17 words
Apparatus as recited in claim 1 wherein said 14 C-labeled substrates are coated on the chamber walls.
Claim 21
Dependent Claim
15 words
Apparatus as recited in claim 1 wherein said 14 C-labeled substrates are solidified in agar.
Claim 22
Dependent Claim
17 words
Apparatus as recited in claim 1 wherein 14 C-labeled substrates are absorbed on a sheet of paper.
Claim 23
Dependent Claim
19 words
Apparatus as recited in claim 1 further comprising means to subject said cells to conditions conducive to biological activity.
Claim 24
Dependent Claim
27 words
Apparatus as recited in claim 22 wherein said culture medium is selected from the class consisting of peptone broth, dextrose free tryptic soy broth, and nutrient broth.
Claim 25
Dependent Claim
15 words
Apparatus as recited in claim 1 further comprising a culture medium disposed in each chamber.
Claim 26
Independent Claim
107 words
A method for characterization of a biologically active agent comprising the steps of: a. providing a plurality of sterile containers with different 14 C-labeled fermentable substrates therein; b. introducing a sample of an axenic biologically active material to be characterized into each container; c. sealing said containers with a gas-tight transparent cover having a carbon dioxide absorber and a scintillation compound coated on the inside thereof in intimate proximity to each other; d. subjecting the sealed containers to conditions conducive to biological activity for a predetermined period of time; and e. thereafter measuring the light emitted by said scintillation compound in each container through the transparent cover.
Claim 27
Independent Claim
107 words
A method for characterization of a biologically active agent comprising the steps of: a. providing a plurality of sterile containers with different 14 C-labeled fermentable substrates therein; b. introducing a sample of an axenic biologically active material to be characterized into each container; c. sealing said containers with a gas-tight transparent cover having a carbon dioxide absorber and a scintillation compound coated on the inside thereof in intimate proximity to each other; d. subjecting the sealed containers to conditions conducive to biological activity for a predetermined period of time; and e. thereafter measuring the light emitted by said scintillation compound in each container through the transparent cover.
Claim 28
Dependent Claim
17 words
A method as recited in claim 25 wherein said containers are formed in a single integral body.
Claim 29
Dependent Claim
16 words
A method as recited in claim 25 wherein said cover comprises a sheet of transparent glass.
Claim 30
Dependent Claim
37 words
A method as recited in claim 25 wherein fermentable substrates are selected from the class consisting of monosaccharides, disaccharides and polysaccharides, amino acids, urea, uracil, carboxylic acids, dicarboxylic acids and salts thereof, hydroxycarboxylic acids, and polyhydroxy alcohols.
Claim 31
Dependent Claim
19 words
A method as recited in claim 25 wherein said fermentable substrates are coated on the walls of the chamber.
Claim 32
Dependent Claim
15 words
A method as recited in claim 25 wherein said fermentable substrates are solidified in agar.
Claim 33
Dependent Claim
17 words
A method as recited in claim 25 wherein said fermentable substrates are absorbed on a paper sheet.
Claim 34
Dependent Claim
14 words
A method as recited in claim 25 wherein said containers contain a culture medium.
Claim 35
Dependent Claim
28 words
A method as recited in claim 32 wherein said culture medium is selected from the class consisting of peptone broth, dextrose free tryptic soy broth, and nutrient broth.
Claim 36
Dependent Claim
35 words
A method as recited in claim 25 wherein said axenic sample of a biologically active agent comprises a viable inoculum of from about 10 5 to about 10 8 microorganisms suspended in a culture medium.
Claim 37
Dependent Claim
58 words
A method as recited in claim 25 wherein said conditions conducive to biological activity comprise incubation in the presence of oxygen at a temperature lying in the range from about 35° to about 37° C at a pH between 6 and 8 for a time interval lying in the range from about 15 minutes to about 3 hours.
Claim 38
Dependent Claim
30 words
A method as recited in claim 35 wherein said predetermined incubation is carried out for a time interval lying in the range from about 1 hour to about 2 hours.
Claim 39
Dependent Claim
18 words
A method as recited in claim 25 wherein said emitted light is measured by means of a photomultiplier.
Claim 40
Dependent Claim
20 words
A method as recited in claim 37 wherein said photomultiplier is successively aligned with the transparent cover of each chamber.
Claim 41
Dependent Claim
22 words
A method as recited in claim 25 wherein said carbon dioxide absorber comprises an alkali metal or an alkaline earth metal hydroxide.
Claim 42
Dependent Claim
15 words
A method as recited in claim 39 wherein said carbon dioxide absorber comprises sodium hydroxide.
Claim 43
Dependent Claim
14 words
A method as recited in claim 25 wherein said scintillation compound comprises 2-, 5-diphenyloxazole.
Claim 44
Dependent Claim
27 words
A method as recited in claim 25 further comprising the step of recording the measured valves for the light emitted by the scintillation compound in each chamber.
Claim 45
Dependent Claim
49 words
A method for characterization of a biologically active agent as recited in claim 25 wherein said biologically active material is an unknown agent, and further comprising the step of comparing the activity pattern of said biologically active material to known activity data in order to identify the unknown agent.
Claim 46
Independent Claim
113 words
A method for characterization of a biologically active agent comprising the steps of: a. providing a plurality of gas-tight cells each having light-permeable wall section and containing: 1. different 14 C-labeled fermentable substrates, 2. a carbon dioxide absorber and a scintillation compound disposed in intimate proximity to each other adjacent said light-permeable wall section and out of contact with said 14 C-labeled fermentable substrates, and 3. an axenic sample of material to be tested for biological activity, b. subjecting said chambers to conditions conductive to biological activity for a predetermined period of time; and c. thereafter measuring the light emitted by said scintillation compound in each chamber through said light permeable wall section.
Claim 47
Independent Claim
113 words
A method for characterization of a biologically active agent comprising the steps of: a. providing a plurality of gas-tight cells each having light-permeable wall section and containing: 1. different 14 C-labeled fermentable substrates, 2. a carbon dioxide absorber and a scintillation compound disposed in intimate proximity to each other adjacent said light-permeable wall section and out of contact with said 14 C-labeled fermentable substrates, and 3. an axenic sample of material to be tested for biological activity, b. subjecting said chambers to conditions conductive to biological activity for a predetermined period of time; and c. thereafter measuring the light emitted by said scintillation compound in each chamber through said light permeable wall section.
Claim 48
Dependent Claim
49 words
A method for characterization of a biologically active agent as recited in claim 44 wherein said biologically active material is an unknown agent, and further comprising the step of comparing the activity pattern of said biologically active material to known activity data in order to identify the unknown agent.
Claim 49
Independent Claim
113 words
A method for characterization of biologically active agent comprising the steps of: a. providing a plurality of sterile containers with different culture media each containing a 14 C-labeled fermentable substance therein; b. introducing a sample of an axenic biologically active material to be characterized into each container; c. sealing said containers with a gas-tight light permeable cover having a carbon dioxide absorber and a scintillation compound coated on the inside thereof in intimate proximity to each other; d. subjecting the sealed containers to conditions conducive to biological activity for a predetermined period to time; and e. thereafter measuring the light emitted by said scintillation compound in each container through the light permeable cover.
Claim 50
Independent Claim
113 words
A method for characterization of biologically active agent comprising the steps of: a. providing a plurality of sterile containers with different culture media each containing a 14 C-labeled fermentable substance therein; b. introducing a sample of an axenic biologically active material to be characterized into each container; c. sealing said containers with a gas-tight light permeable cover having a carbon dioxide absorber and a scintillation compound coated on the inside thereof in intimate proximity to each other; d. subjecting the sealed containers to conditions conducive to biological activity for a predetermined period to time; and e. thereafter measuring the light emitted by said scintillation compound in each container through the light permeable cover.
Claim 51
Dependent Claim
49 words
A method for characterization of a biologically active agent as recited in claim 46 wherein said biologically active material is an unknown agent, and further comprising the step of comparing the activity pattern of said biologically active material to known activity data in order to identify the unknown agent.
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) | 1979-12-14 | 1980-06-14 | 1980-12-14 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1983-12-14 | 1984-06-14 | 1984-12-14 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1987-12-14 | 1988-06-14 | 1988-12-14 | $7700.00 | Window Closed / Lapsed |