Methods for detecting antibody that specifically bind a siderophore receptor polypeptide
The present invention provides compositions including siderophore receptor polypeptides and porins from gram negative microbes, and preferably, lipopolysaccharide 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.
1. A method for detecting antibody comprising:
contacting antibody with a preparation to result in a mixture, wherein the preparation comprises at least two siderophore receptor polypeptides isolated from a gram negative microbe and lipopolysaccharide at a concentration of no greater than about 10.0 endotoxin units per milliliter (EU/ml), wherein the at least two siderophore receptor polypeptides have molecular weights of between about 60 kDa and about 100 kDa as determined by separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis;
incubating the mixture under conditions to allow the antibody to specifically bind a siderophore receptor polypeptide to form a siderophore receptor polypeptide:antibody complex; and
detecting the siderophore receptor polypeptide:antibody complex, wherein detection of the siderophore receptor polypeptide:antibody complex indicates the presence of antibody that specifically binds a siderophore receptor polypeptide.
2. The method of claim 1 wherein the antibody is present in a biological sample from an animal, and wherein the presence of antibody that specifically binds a siderophore receptor polypeptide indicates the animal may have a condition caused by a microbe expressing siderophore receptor polypeptides.
3. The method of claim 2 wherein the biological sample comprises blood, milk, or colostrum.
4. The method of claim 1 wherein the antibody is present in a biological sample from an animal, wherein the animal has been immunized by administration of a composition comprising at least 2 siderophore receptor polypeptides, and wherein the presence of antibody that specifically binds a siderophore receptor polypeptide indicates the animal has a serological response to the administration.
5. The method of claim 4 wherein the biological sample comprises blood, milk, or colostrum.
6. The method of claim 1 wherein the preparation comprises a solid matrix to which the siderophore receptor polypeptides are affixed.
7. The method of claim 1 wherein the detecting comprises a radioimmunoassay or an enzyme-linked immunoassay.
8. The method of claim 2 wherein the animal is an avian, a bovine, a caprine, a porcine, an ovine, a Bison, a companion animal, a member of the family Ceridae, or a human.
9. The method of claim 4 wherein the animal is an avian, a bovine, a caprine, a porcine, an ovine, a Bison, a companion animal, a member of the family Ceridae, or a human.