Polypeptides obtained from gram negative microbes
The present invention provides compositions including at least two siderophore receptor polypeptides and at least two porins from a gram negative microbe, and preferably, lipopolysaccharide at a concentration of no greater than about 10.0 endotoxin units per milliliter. The present invention also provides methods of makino and methods of using such compositions.
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 as determined by separation by sodium dodecyl-polyacrylamide gel electrophoresis.
8. The composition of claim 1 wherein the at least two porins have molecular weights of between about 30 kDa and about 43 kDa as determined by separation by sodium dodecyl-polyacrylamide gel electrophoresis.
9. The composition of claim 6 wherein the gram negative microbe is Salmonella spp., and wherein each of the at least two SRPs have a molecular weight selected from the group consisting of between about 87 kDa and about 91 kDa, between about 82 kDa and about 86 kDa, and between about 69 kDa and about 75 kDa, wherein the molecular weight is determined by separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
10. A first 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), wherein the concentration of LPS is no greater than the concentration of LPS in a second composition comprising:
the at least two siderophore receptor polypeptides isolated from the gram negative microbe;
the at least two porins isolated from the gram negative microbe; and
lipopolysaccharide;
wherein the second composition is produced by a process comprising:
providing the gram negative microbe;
disrupting the gram negative microbe in a buffer;
solubilizing the disrupted gram negative microbe for greater than about 24 hours in a solution comprising sarcosine to result in solubilized and insoluble cellular material, 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 the at least two SRPs, the at least two porins, and LPS.
11. The first composition of claim 10 further comprising a pharmaceutically acceptable carrier.
12. The first composition of claim 10 wherein the LPS concentration is no greater than about 10.0 EU/ml.
13. The first composition of claim 10 wherein the gram negative microbe is an enteropathogen.
14. The first composition of claim 10 wherein the gram negative microbe is a member of the family Enterobacteriaceae.
15. The first composition of claim 10 wherein the gram negative microbe is a member of the tribe Escherichieae or Salmonelleae.
16. The first composition of claim 10 wherein the gram negative microbe is Salmonella spp. or Escherichia coli.
17. The first composition of claim 10 wherein the at least two SRPs have molecular weights of between about 60 kDa and about 100 kDa as determined by separation by sodium dodecyl-polyacrylamide gel electrophoresis.
18. The first composition of claim 10 wherein the at least two porins have molecular weights of between about 30 kDa and about 43 kDa as determined by separation by sodium dodecyl-polyacrylamide gel electrophoresis.
19. The first composition of claim 16 wherein the gram negative microbe is Salmonella spp., and wherein each of the at least two SRPs have a molecular weight selected from the group consisting of between about 87 kDa and about 91 kDa, between about 82 kDa and about 86 kDa, and between about 69 kDa and about 75 kDa, wherein the molecular weight is determined by separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.