US2003036639A1PendingUtilityA1

Immunizing compositions and methods of use

57
Priority: Jan 3, 2001Filed: Jan 3, 2002Published: Feb 20, 2003
Est. expiryJan 3, 2021(expired)· nominal 20-yr term from priority
Inventors:Daryll A. EmeryDarren E. StraubDonavan E. ZammertGayla K. Kallevig
A61K 31/739Y10S424/826A61K 39/39Y10S424/827A61P 31/04A61K 39/0275Y10S424/829Y10S424/823A61K 2039/552A61K 39/085A61K 2039/55572Y10S424/825A61P 37/02Y10S424/828C07K 14/24A61P 37/04Y10S424/824G01N 33/53Y02A50/30
57
PatentIndex Score
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References
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Claims

Abstract

The present invention provides compositions including siderophore receptor polypeptides and porins from gram negative microbes, and preferably, lipopolysaccarhide at a concentration of no greater than about 10.0 endotoxin units per milliliter. The present invention also provides methods of making and methods of using such compositions.

Claims

exact text as granted — not AI-modified
What is claimed is:  
     
         1 . A composition comprising: 
 at least two siderophore receptor polypeptides (SRPs) isolated from a gram negative microbe;    at least two porins isolated from the gram negative microbe; and    lipopolysaccharide (LPS) at a concentration of no greater than about 10.0 endotoxin units per milliliter (EU/ml).    
     
     
         2 . The composition of  claim 1  further comprising a pharmaceutically acceptable carrier.  
     
     
         3 . The composition of  claim 1  wherein the gram negative microbe is an enteropathogen.  
     
     
         4 . The composition of  claim 1  wherein the gram negative microbe is a member of the family Enterobacteriaceae.  
     
     
         5 . The composition of  claim 1  wherein the gram negative microbe is a member of the tribe Escherichieae or Salmonelleae.  
     
     
         6 . The composition of  claim 1  wherein the gram negative microbe is Salmonella spp. or  Escherichia coli.    
     
     
         7 . The composition of  claim 1  wherein the at least two SRPs have molecular weights of between about 60 kDa and about 100 kDa.  
     
     
         8 . The composition of  claim 1  wherein the at least two porins have molecular weights of between about 30 kDa and about 43 kDa.  
     
     
         9 . A method for inducing the production of antibody in an animal, the method comprising administering to an animal an effective amount of a composition comprising: 
 at least two SRPs isolated from a gram negative microbe;    at least two porins isolated from the gram negative microbe;    LPS at a concentration of no greater than about 10.0 EU/ml; and    a pharmaceutically acceptable carrier, wherein the composition induces in the animal antibody that specifically binds at least one SRP or at least one porin.    
     
     
         10 . The method of  claim 9  wherein the gram negative microbe is an enteropathogen.  
     
     
         11 . The method of  claim 9  wherein the gram negative microbe is a member of the family Enterobacteriaceae.  
     
     
         12 . The method of  claim 9  wherein the gram negative microbe is a member of the tribe Escherichieae or Salmonelleae.  
     
     
         13 . The method of  claim 9  wherein the gram negative microbe is Salmonella spp. or  Escherichia coli.    
     
     
         14 . The method of  claim 9  wherein the at least two SRPs have molecular weights of between about 60 kDa and about 100 kDa.  
     
     
         15 . The method of  claim 9  wherein the at least two porins have molecular weights of between about 30 kDa and about 43 kDa.  
     
     
         16 . The method of  claim 9  wherein the animal is an avian, a bovine, a caprine, a porcine, or an ovine.  
     
     
         17 . The method of  claim 9  wherein the animal is a bovine, and wherein the bovine exhibits a phenotype selected from the group consisting of decreased somatic cell count, increased milk production, decreased fecal shedding, and increased weight.  
     
     
         18 . A method for inducing the production of antibody in an animal, the method comprising administering to an animal an effective amount of a composition comprising: 
 at least four SRPs isolated from a gram positive microbe; and    a pharmaceutically acceptable carrier, wherein the composition induces in the animal antibody to the SRP.    
     
     
         19 . The method of  claim 18  wherein the gram positive microbe is a member of the family Micrococcaceae.  
     
     
         20 . The method of  claim 18  wherein the gram positive microbe is  Staphylococcus aureus.    
     
     
         21 . The method of  claim 18  wherein the at least four SRPs have molecular weights of between about 60 kDa and about 100 kDa.  
     
     
         22 . A method for treating an animal for a high somatic cell count, the method comprising administering to a milk producing animal having or at risk of having a high somatic cell count an effective amount of a composition comprising: 
 at least two SRPs isolated from a gram negative microbe;    at least two porins isolated from the gram negative microbe;    LPS at a concentration of no greater than about 10.0 EU/ml; and    a pharmaceutically acceptable carrier.    
     
     
         23 . The method of  claim 22  wherein the gram negative microbe is an enteropathogen.  
     
     
         24 . The method of  claim 22  wherein the gram negative microbe is a member of the family Enterobacteriaceae.  
     
     
         25 . The method of  claim 22  wherein the gram negative microbe is a member of the tribe Escherichieae or Salmonelleae.  
     
     
         26 . The method of  claim 22  wherein the gram negative microbe is Salmonella spp. or  Escherichia coli.    
     
     
         27 . The composition of  claim 22  wherein the at least two SRPs have molecular weights of between about 60 kDa and about 100 kDa.  
     
     
         28 . The composition of  claim 22  wherein the at least two porins have molecular weights of between about 30 kDa and about 43 kDa.  
     
     
         29 . The method of  claim 22  wherein the animal is a bovine.  
     
     
         30 . A method for reducing fecal shedding of a microbe in an animal's intestinal tract, the method comprising administering to an animal an effective amount of a composition comprising: 
 at least two SRPs isolated from a gram negative microbe;    at least two porins isolated from the gram negative microbe;    LPS at a concentration of no greater than about 10.0 EU/ml; and    a pharmaceutically acceptable carrier.    
     
     
         31 . The method of  claim 30  wherein the gram negative microbe is an enteropathogen.  
     
     
         32 . The method of  claim 30  wherein the gram negative microbe is a member of the family Enterobacteriaceae.  
     
     
         33 . The method of  claim 30  wherein the gram negative microbe is a member of the tribe Escherichieae or Salmonelleae.  
     
     
         34 . The method of  claim 30  wherein the gram negative microbe is Salmonella spp. or  Escherichia coli.    
     
     
         35 . The method of  claim 30  wherein the at least two SRPs have molecular weights of between about 60 kDa and about 100 kDa.  
     
     
         36 . The method of  claim 30  wherein the at least two porins have molecular weights of between about 30 kDa and about 43 kDa.  
     
     
         37 . The method of  claim 30  wherein the animal is an avian, a bovine, a caprine, a porcine, or an ovine.  
     
     
         38 . The method of  claim 30  wherein the microbe present in the animal's intestinal tract is a gram negative microbe or a gram positive microbe.  
     
     
         39 . A method for treating an animal for low milk production, the method comprising administering to a milk producing animal having or at risk of having a low milk production an effective amount of a composition comprising: 
 at least two SRPs isolated from a gram negative microbe;    at least two porins isolated from the gram negative microbe;    LPS at a concentration of no greater than about 10.0 EU/ml; and    a pharmaceutically acceptable carrier.    
     
     
         40 . The method of  claim 39  wherein the gram negative microbe is an enteropathogen.  
     
     
         41 . The method of  claim 39  wherein the gram negative microbe is a member of the family Enterobacteriaceae.  
     
     
         42 . The method of  claim 39  wherein the gram negative microbe is a member of the tribe Escherichieae or Salmonelleae.  
     
     
         43 . The method of  claim 39  wherein the gram negative microbe is Salmonella spp. or  Escherichia coli.    
     
     
         44 . The method of  claim 39  wherein the at least two SRPs have molecular weights of between about 60 kDa and about 100 kDa.  
     
     
         45 . The method of  claim 39  wherein the at least two porins have molecular weights of between about 30 kDa and about 43 kDa.  
     
     
         46 . The method of  claim 39  wherein the animal is a bovine.  
     
     
         47 . A method for treating mastitis in a milk producing animal, the method comprising administering to a milk producing animal having or at risk of having mastitis an effective amount of a composition comprising: 
 at least two SRPs isolated from a gram negative microbe;    at least two porins isolated from the gram negative microbe;    LPS at a concentration of no greater than about 10.0 EU/ml; and    a pharmaceutically acceptable carrier.    
     
     
         48 . The method of  claim 47  wherein the gram negative microbe is an enteropathogen.  
     
     
         49 . The method of  claim 47  wherein the gram negative microbe is a member of the family Enterobacteriaceae.  
     
     
         50 . The method of  claim 47  wherein the gram negative microbe is a member of the tribe Escherichieae or Salmonelleae.  
     
     
         51 . The method of  claim 47  wherein the gram negative microbe is Salmonella spp. or  Escherichia coli.    
     
     
         52 . The method of  claim 47  wherein the at least two SRPs have molecular weights of between about 60 kDa and about 100 kDa.  
     
     
         53 . The method of  claim 47  wherein the at least two porins have molecular weights of between about 30 kDa and about 43 kDa  
     
     
         54 . The method of  claim 47  wherein the animal is a bovine.  
     
     
         55 . A method for treating metritis in an animal, the method comprising administering to a milk producing animal having or at risk of having metritis an effective amount of a composition comprising: 
 at least two SRPs isolated from a gram negative microbe;    at least two porins isolated from the gram negative microbe;    LPS at a concentration of no greater than about 10.0 EU/ml; and    a pharmaceutically acceptable carrier.    
     
     
         56 . The method of  claim 55  wherein the gram negative microbe is an enteropathogen.  
     
     
         57 . The method of  claim 55  wherein the gram negative microbe is a member of the family Enterobacteriaceae.  
     
     
         58 . The method of  claim 55  wherein the gram negative microbe is a member of the tribe Escherichieae or Salmonelleae.  
     
     
         59 . The method of  claim 55  wherein the gram negative microbe is Salmonella spp. or  Escherichia coli.    
     
     
         60 . The method of  claim 55  wherein the at least two SRPs have molecular weights of between about 60 kDa and about 100 kDa.  
     
     
         61 . The method of  claim 55  wherein the at least two porins have molecular weights of between about 30 kDa and about 43 kDa  
     
     
         62 . The method of  claim 55  wherein the animal is a bovine.  
     
     
         63 . A method for isolating outer membrane polypeptides, the method comprising 
 providing a gram negative microbe;    disrupting the gram negative microbe in a buffer;    solubilizing the disrupted gram negative microbe; and    isolating molecules of the gram negative microbe, wherein the isolated molecules comprise outer membrane polypeptides comprising at least two SRPs and at least two porins, and LPS at a concentration of no greater than about 10.0 EU/ml.    
     
     
         64 . A method for isolating outer membrane polypeptides, the method comprising 
 providing a gram negative microbe;    disrupting the gram negative microbe in a buffer, wherein the gram negative microbe is present in the buffer at a concentration of between about 720 grams of microbe per 1,000 milliliters of buffer and about 1,080 grams of microbe per 1,000 milliliters of buffer;    solubilizing the disrupted gram negative microbe for greater than about 24 hours in a solution comprising sarcosine, wherein a ratio of the sarcosine to gram weight of disrupted gram negative microbe is between about 0.8 gram sarcosine per about 4.5 grams of disrupted gram negative microbe and about 1.2 grams sarcosine per about 4.5 grams of disrupted gram negative microbe; and    isolating molecules of the gram negative microbe, wherein the isolated molecules comprise outer membrane polypeptides comprising at least two SRPs and at least two porins.    
     
     
         65 . The method of  claim 64  wherein the isolated molecules further comprise LPS at a concentration of no greater than about 10.0 EU/ml.  
     
     
         66 . A method for isolating outer membrane polypeptides, the method comprising 
 providing a gram negative microbe;    disrupting the gram negative microbe;    solubilizing the disrupted gram negative microbe in a solution comprising sarcosine, wherein ratio of the sarcosine to gram weight of disrupted gram negative microbe is between about 0.8 gram sarcosine per about 4.5 grams of disrupted gram negative microbe and about 1.2 grams sarcosine per about 4.5 grams of disrupted gram negative microbe; and    isolating molecules of the gram negative microbe, wherein the isolated molecules comprise outer membrane polypeptides comprising at least two SRPs and at least two porins.    
     
     
         67 . The method of  claim 66  wherein the isolated molecules further comprise LPS at a concentration of no greater than about 10.0 EU/ml.  
     
     
         68 . A method for isolating outer membrane polypeptides, the method comprising 
 providing a gram negative microbe;    disrupting the gram negative microbe;    solubilizing the disrupted gram negative microbe for greater than about 24 hours; and    isolating molecules of the gram negative microbe, wherein the isolated molecules comprise outer membrane polypeptides comprising at least two SRPs and at least two porins.    
     
     
         69 . The method of  claim 68  wherein the isolated molecules further comprise LPS at a concentration of no greater than about 10.0 EU/ml.  
     
     
         70 . A method for isolating outer membrane polypeptides, the method comprising 
 providing a gram negative microbe;    disrupting the disrupted gram negative microbe in a buffer, wherein the gram negative microbe is present in the buffer at a concentration of between about 720 grams of microbe per 1,000 milliliters of buffer to about 1,080 grams of microbe per 1,000 milliliters of buffer;    solubilizing the gram negative microbe; and    isolating molecules of the gram negative microbe, wherein the isolated molecules comprise outer membrane polypeptides comprising at least two SRPs and at least two porins.    
     
     
         71 . The method of  claim 70  wherein the isolated molecules further comprise LPS at a concentration of no greater than about 10.0 EU/ml.

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