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US4158049A Expired / Lapsed US ⏱️ 0.0 years remaining

Antigen fraction of Schistosoma mansoni eggs suitable for testing for schistosomiasis

Assignee: Clark Edna Mcconnell Foundation
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Filing: 1976-09-27
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Granted: 1979-06-12
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Expires: 1996-09-27
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Abstract

Abstract Schistosoma mansoni eggs are ground to liberate soluble egg antigens which are fractionated and purified by absorption, gel filtration and/or ion exchange chromatography, and then radioactively labelled with 125 I. A fraction, identified as MSA 1 , is specific to the mature egg stage of Schistosoma mansoni and thus can be used to detect the presence of antibody to mature Schistosoma mansoni eggs in a patient's serum even as compared with mature eggs of different Schistosoma (species specific) or of Schistosoma mansoni worms but not mature eggs (stage specific).

Inventors

  • 👤 Ronald P. Pelley
  • 👤 Kenneth S. Warren

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-11-01 10:46
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/5308; A; A61; A61K; A61K39/00; A61K39/0003; Y; Y02; Y02A; Y02A50/00; Y02A50/30; Y10; Y10S; Y10S436/00; Y10S436/811

Claims Analysis

Total Claims: 15
Claim 1
Independent Claim 83 words
A fraction of Schistosoma mansoni egg antigen specific to Schistosoma mansoni mature eggs, and which has the following properties: ______________________________________ Polyacrylamide gel R.F. 0.34 ± 0.01 Staining Protein (Coomassie Blue) + Carbohydrate (Periodic acid, Schiff reagent) + Lipid (Sudan Black) - Concanavalin A Binding Tightly Adsorbed CsCl Buoyant Density 1.43 ± .01 g/cc (homogeneous) MOLECULAR WEIGHT Gel Fitration 148,000 ± 5,000 Velocity Sedimentation 90-100,000 Polyacrylamide gel (R.F.) 50,000 (.50) Charge (Isoelectric Point) pH 3.514 4.5 ______________________________________ said fraction being suitable to detect Schistosomiasis.
Claim 2
Dependent Claim 83 words
A fraction of Schistosoma mansoni egg antigen specific to Schistosoma mansoni mature eggs, and which has the following properties: ______________________________________ Polyacrylamide gel R.F. 0.34 ± 0.01 Staining Protein (Coomassie Blue) + Carbohydrate (Periodic acid, Schiff reagent) + Lipid (Sudan Black) - Concanavalin A Binding Tightly Adsorbed CsCl Buoyant Density 1.43 ± .01 g/cc (homogeneous) MOLECULAR WEIGHT Gel Fitration 148,000 ± 5,000 Velocity Sedimentation 90-100,000 Polyacrylamide gel (R.F.) 50,000 (.50) Charge (Isoelectric Point) pH 3.514 4.5 ______________________________________ said fraction being suitable to detect Schistosomiasis.
Claim 3
Dependent Claim 10 words
A fraction according to claim 1, which is radioactively labeled.
Claim 4
Dependent Claim 13 words
A fraction according to claim 1, which is radioactively labeled with 125 I.
Claim 5
Dependent Claim 73 words
A fraction of Schistosoma mansoni egg antigen which has the following approximate properties: ______________________________________ Polyacrylamide gel R.F. 0.48 ± 0.01 Staining Protein (Coomassie Blue) + Carbohydrate (Periodic acid, Schiff reagent) - Lipid (Sudan Black) - Concanavalin A Binding Loosely adsorbed or not adsorbed at all CsCl Buoyant Density 1.53 g/cc.sup.2 MOLECULAR WEIGHT Gel Filtration 80,000 ± 5,00 Velocity Sedimentation 40,000 Polyacrylamide gel (R.F.) 69,000 (.33) ______________________________________ said fraction being suitable to detect Schistosomiasis.
Claim 6
Dependent Claim 73 words
A fraction of Schistosoma mansoni egg antigen which has the following approximate properties: ______________________________________ Polyacrylamide gel R.F. 0.48 ± 0.01 Staining Protein (Coomassie Blue) + Carbohydrate (Periodic acid, Schiff reagent) - Lipid (Sudan Black) - Concanavalin A Binding Loosely adsorbed or not adsorbed at all CsCl Buoyant Density 1.53 g/cc.sup.2 MOLECULAR WEIGHT Gel Filtration 80,000 ± 5,00 Velocity Sedimentation 40,000 Polyacrylamide gel (R.F.) 69,000 (.33) ______________________________________ said fraction being suitable to detect Schistosomiasis.
Claim 7
Dependent Claim 87 words
A fraction of Schistosoma mansoni egg antigen which has the following approximate properties: ______________________________________ Predominant Occasionally Polyacrylamide gel R.F. species observed species ______________________________________ Staining 0.22 ± .01 0.08 0.18 Protein (Coomassie Blue) + + + Carbohydrate (Per- iodic acid, Schiff reagent) + + + Lipid + + + Concanavalin A Binding Loosely adsorbed CsCl Buoyant Density Heterogeneous N.D. N.D. (1.51-1.37) MOLECULAR WEIGHT Gel Filtration 450,000±8,000 N.D. N.D. Velocity Sedimenta- tion 200,000 N.D. N.D. Polyacrylamide gel (R.F.) 200,000 N.D. N.D. ______________________________________ said fraction being suitable to detect Schistosomiasis.
Claim 8
Dependent Claim 87 words
A fraction of Schistosoma mansoni egg antigen which has the following approximate properties: ______________________________________ Predominant Occasionally Polyacrylamide gel R.F. species observed species ______________________________________ Staining 0.22 ± .01 0.08 0.18 Protein (Coomassie Blue) + + + Carbohydrate (Per- iodic acid, Schiff reagent) + + + Lipid + + + Concanavalin A Binding Loosely adsorbed CsCl Buoyant Density Heterogeneous N.D. N.D. (1.51-1.37) MOLECULAR WEIGHT Gel Filtration 450,000±8,000 N.D. N.D. Velocity Sedimenta- tion 200,000 N.D. N.D. Polyacrylamide gel (R.F.) 200,000 N.D. N.D. ______________________________________ said fraction being suitable to detect Schistosomiasis.
Claim 9
Dependent Claim 77 words
The process for fractionating Schistosoma mansoni egg antigen comprising grinding Schistosoma mansoni eggs in water to form a solution thereof, subjecting said solution to affinity chromatography and eluting to form a third fraction which was most lightly held, a second fraction which was less lightly held and a first fraction which was strongly held, and purifying at least said first fraction, said first fraction containing antigens as defined in claim 1 specific to Schistosoma mansoni mature eggs.
Claim 10
Dependent Claim 42 words
The process of claim 6, wherein said affinity chromatography is effected on a galactan, elution of said first fraction is effected by at least one of α-methyl mannoside or α-methyl glucoside, and including the further step of radioactively labeling said first fraction.
Claim 11
Dependent Claim 14 words
The process of claim 7, wherein the radioactive labeling is effected with 125 I.
Claim 12
Dependent Claim 50 words
The process of claim 6, wherein purification of at least the first fraction is effected by ion exchange chromatography using a salt eluant, any third fraction material present being most lightly held, any second fraction material present being less lightly held, and the first fraction material being most strongly held.
Claim 13
Dependent Claim 76 words
The process of claim 8, wherein purification of at least the first fraction is effected by ion exchange chromatography using a salt eluant, any third fraction material present being most lightly held, any second fraction material being less lightly held, and the first fraction material being most strongly held, and further including desalting said first fraction at some stage subsequent to radioactive labeling to remove at least part of any salts introduced during said radioactive labeling.
Claim 14
Dependent Claim 63 words
A process for determining the presence of antibodies to mature schistosome eggs in a patient's serum comprising mixing said serum with a radioactively labeled fraction produced by the process of claim 7, separating any precipitate which forms, and measuring the radioactivity of said precipitate, the level of radioactivity being an index of the amount of antibodies to mature schistosome eggs in said serum.
Claim 15
Dependent Claim 61 words
A process for determining the presence of antibodies to mature Schistosoma mansoni eggs in a patient's serum comprising mixing said serum with a radioactively labeled fraction according to claim 2, separating any precipitate which forms, and measuring the radioactivity of said precipitate, the level of radioactivity being an index of the amount of antibodies to Schistosoma mansoni eggs in said serum.

References Cited (Backward Citations)

US3860483A Cited

USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1982-06-11 1982-12-11 1983-06-12 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1986-06-11 1986-12-11 1987-06-12 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1990-06-11 1990-12-11 1991-06-12 $7700.00 Window Closed / Lapsed

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