US4298597A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Vaccine for diarrhea caused by E. coli
Assignee: University of Saskatchewan
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Filing: 1979-09-04
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Granted: 1981-11-03
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Expires: 1999-09-04
Abstract
Abstract A relatively safe and effective vaccine for coliform diarrhea and related infections is prepared from semipurified K99 and other concentrated subcellular antigens and organelles of selected. E. coli strains. For this purpose broth cultures of E. coli bacteria are mechanically sheared or sonicated to disrupt the bacterial cells, then centrifuged and the supernatent containing subcellular particles and components is concentrated by ultrafiltration and sterilized by the addition of a preservative and/or stabilizer.
Inventors
- 👤 Stephen D. Acres
- 👤 Robert A. Kapitany
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-06 09:13
CPC Classifications
A; A61; A61K; A61K39/00; A61K39/02; A61K39/025; A61K39/0258; Y; Y02; Y02A; Y02A50/00; Y02A50/30; Y10; Y10S; Y10S424/00; Y10S424/823
Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
151 words
A vaccine for promoting in cattle the formation of antibodies against antigens from enterotoxigenic E. coli comprising an aqueous suspension or solution containing an effective amount of K99 antigen, polysaccharide K antigens, lipopolysaccharide O antigens, enterotoxins, or combinations of the foregoing constituents, said vaccine lacking whole intact cells, but containing cell fragments and other cellular and culture broth constituents having a molecular weight greater than 1000; said vaccine being prepared by a method comprising: (a) growing several selected enterotoxigenic strains of E. coli in a broth culture medium under aerobic conditions, with agitation; (b) pooling the E. coli cultures; (c) disrupting the bacteria by input of energy for removal of cell wall polysaccharide outer layers and pili; (d) removing intact cells from the disrupted suspension; and (e) selectively concentrating the subcellular antigens for removal of undesired low molecular weight components and excess water by passing the suspension through an ultrafiltration membrane.
Claim 2
Dependent Claim
151 words
A vaccine for promoting in cattle the formation of antibodies against antigens from enterotoxigenic E. coli comprising an aqueous suspension or solution containing an effective amount of K99 antigen, polysaccharide K antigens, lipopolysaccharide O antigens, enterotoxins, or combinations of the foregoing constituents, said vaccine lacking whole intact cells, but containing cell fragments and other cellular and culture broth constituents having a molecular weight greater than 1000; said vaccine being prepared by a method comprising: (a) growing several selected enterotoxigenic strains of E. coli in a broth culture medium under aerobic conditions, with agitation; (b) pooling the E. coli cultures; (c) disrupting the bacteria by input of energy for removal of cell wall polysaccharide outer layers and pili; (d) removing intact cells from the disrupted suspension; and (e) selectively concentrating the subcellular antigens for removal of undesired low molecular weight components and excess water by passing the suspension through an ultrafiltration membrane.
Claim 3
Dependent Claim
20 words
A vaccine as in claim 1 wherein the cellular and culture broth constituents have a molecular weight greater than 5,000.
Claim 4
Dependent Claim
20 words
A vaccine as in claim 1 wherein the cellular and culture broth constituents have a molecular weight greater than 10,000.
Claim 5
Dependent Claim
21 words
A vaccine as in claim 1 wherein the aqueous suspension or solution contains 0.1 μg/ml (micrograms/ml) to 2 mg/ml K99 antigen.
Claim 6
Dependent Claim
19 words
A vaccine as in claim 1 wherein the aqueous suspension or solution contains 0.5 to 500 μg/ml K99 antigen.
Claim 7
Dependent Claim
20 words
A vaccine as in claim 1 wherein the aqueous suspension or solution contains 20 to 320 ng/ml (nanograms/ml) heat-stable enterotoxins.
Claim 8
Dependent Claim
19 words
A vaccine as in claim 1 wherein the aqueous suspension or solution contains 30 to 100 ng/ml heat-stable enterotoxins.
Claim 9
Dependent Claim
37 words
A vaccine as in claim 1 wherein the aqueous suspension or solution contains a combination of constituents selected from at least two of the groups consisting of K99 antigen, polysaccharide K antigens, lipopolysaccharide O antigens, and enterotoxins.
Claim 10
Dependent Claim
15 words
A vaccine as in claim 8 wherein one of the selected constituents is K99 antigen.
Claim 11
Dependent Claim
29 words
A vaccine as in claim 9 wherein said combination is K99 antigen, at least one polysaccharide K antigen, at least one lipopolysaccharide O antigen, and at least one enterotoxin.
Claim 12
Dependent Claim
52 words
A vaccine as in claim 1 wherein said polysaccharide K antigens are selected from the group consisting of K25, K28, K30, K35, K85, K?, and mixtures thereof; said lipopolysaccharide O antigens are selected from the group consisting of 08, 09, 020, 0101, and mixtures thereof; and said enterotoxins are heat stable enterotoxins.
Claim 13
Independent Claim
177 words
A method of preventing neonatal diarrhea in calves comprising the step of administering to the pregnant cow prior to parturition a vaccine containing an effective amount of K99 antigen, polysaccharide K antigens, lipopolysaccharide O antigens, enterotoxins, or combinations of the foregoing constituents, said vaccine lacking whole intact cells, but containing cell fragments and other cellular and broth constituents having a molecular weight greater than 1000, in combination with a suitable adjuvant, said effective amount of vaccine inducing in the cow effective colostral levels of antibodies against said antigens from enterotoxigenic E. coli; said vaccine being prepared by a method comprising: (a) growing several selected enterotoxigenic strains of E. coli in a broth culture medium under aerobic conditions, with agitation; (b) pooling the E. coli cultures; (c) disrupting the bacteria by input of energy for removal of cell wall polysaccharide outer layers and pili; (d) removing intact cells from the disrupted suspension; and (e) selectively concentrating the subcellular antigens for removal of undesired low molecular weight components and excess water by passing the suspension through an ultrafiltration membrane.
Claim 14
Independent Claim
177 words
A method of preventing neonatal diarrhea in calves comprising the step of administering to the pregnant cow prior to parturition a vaccine containing an effective amount of K99 antigen, polysaccharide K antigens, lipopolysaccharide O antigens, enterotoxins, or combinations of the foregoing constituents, said vaccine lacking whole intact cells, but containing cell fragments and other cellular and broth constituents having a molecular weight greater than 1000, in combination with a suitable adjuvant, said effective amount of vaccine inducing in the cow effective colostral levels of antibodies against said antigens from enterotoxigenic E. coli; said vaccine being prepared by a method comprising: (a) growing several selected enterotoxigenic strains of E. coli in a broth culture medium under aerobic conditions, with agitation; (b) pooling the E. coli cultures; (c) disrupting the bacteria by input of energy for removal of cell wall polysaccharide outer layers and pili; (d) removing intact cells from the disrupted suspension; and (e) selectively concentrating the subcellular antigens for removal of undesired low molecular weight components and excess water by passing the suspension through an ultrafiltration membrane.
Claim 15
Dependent Claim
21 words
A method as in claim 12, wherein the vaccine is administered to the cow at least a week prior to parturition.
Claim 16
Dependent Claim
25 words
A method as in claim 12, wherein the vaccine is administered to the cow about 6 weeks and again about 3 weeks prior to parturition.
Claim 17
Dependent Claim
23 words
A method as in claim 12, wherein the vaccine administered to the cow contains 0.5 μg to 10 mg K99 antigen per dose.
Claim 18
Dependent Claim
23 words
A method as in claim 12, wherein the vaccine administered to the cow contains 2.5 μg to 2.5 mg K99 antigen per dose.
Claim 19
Dependent Claim
39 words
A method as in claim 12, wherein the effective serum antibody titers induced in the cow are as follows: against K99, from 1/8 to 1/2048 against polysaccharide K, from 1/2 to 1/128 against lipopolysaccharide O, from 1/16 to 1/128
Claim 20
Dependent Claim
30 words
A method as in claim 12, wherein the minimum serum antibody titers induced in the cow are as follows: against K99, 1/2 against polysaccharide K, 1/2 against lipopolysaccharide O, 1/8
Claim 21
Dependent Claim
34 words
A method as in claim 12, wherein the effective colostral antibody titers induced are: against K99, from 1/8 to 1/8192 against polysaccharide K, from 1/4 to 1/128 against lipopolysaccharide O, from 1/32 to 1/1024
Claim 22
Dependent Claim
27 words
A method as in claim 12, wherein the minimum colostral antibody titers induced are as follows: against K99, 1/8 against polysaccharide K, 1/8 against lipopolysaccharide O, 1/8
Claim 23
Dependent Claim
23 words
A method as in claim 12, wherein the adjuvant is selected from the group of Freund's Adjuvant, aluminum hydroxide and aluminum potassium sulfate.
References Cited (Backward Citations)
US3907987A
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1984-11-02 | 1985-05-04 | 1985-11-03 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1988-11-02 | 1989-05-04 | 1989-11-03 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1992-11-02 | 1993-05-04 | 1993-11-03 | $7700.00 | Window Closed / Lapsed |