Recombinant VapA and VapC peptides and uses thereof
The present invention provides a recombinant protein comprising consecutive amino acids, the sequence of which is substantially identical to a sequence of amino acids present in a Rhodococcus equi virulence-associated protein and compositions containing fusion proteins of the invention. The present invention also provides uses of the compositions in the manufacture of hyperimmune plasma against Rhodococcus equi , in producing a hyperimmune plasma against Rhodococcus equi in protecting an animal against Rhodococcus equi and in protecting a newborn animal against Rhodococcus equi.
1. An isolated recombinant protein consisting of:
(a) consecutive amino acids 29-189 of Rhodococcus equi virulence-associated protein A (VapA), (i) the sequence of which is set forth in SEQ ID NO:1, or (ii) varies from such sequence by not more than five amino acids; or consecutive amino acids 29-173 of Rhodococcus equi virulence-associated protein (VapC), the sequence of which is set forth in SEQ ID NO:3, or (ii) varies from such sequence by not more than five amino acids,
(b) consecutive amino acids which consist of a 6-His tag, a poly-6-His tag, or a glutathione S-transferase tag, and
(c) a linker sequence between (a) and (b).
2. The isolated recombinant protein of claim 1 , wherein the sequence of the isolated recombinant protein is the sequence set forth in SEQ ID NO: 18.
3. The isolated recombinant protein of claim 1 , wherein the sequence of the isolated recombinant protein is the sequence set forth in SEQ ID NO: 19.
4. The isolated recombinant protein of claim 1 , wherein the sequence of the isolated recombinant protein is the sequence set forth in SEQ ID NO: 20.
5. The isolated recombinant protein of claim 1 , wherein the sequence of the isolated recombinant protein is the sequence set forth in SEQ ID NO: 21.
6. A composition comprising the isolated recombinant protein of claim 1 and a carrier.
7. The composition of claim 6 , wherein the carrier is an aqueous or non-aqueous solution, suspension or emulsion and the recombinant protein is present in an amount of about 0.25 mg/ml to about 2.5 mg/ml.
8. The composition of claim 7 , wherein the recombinant protein is present in an amount of about 0.5 mg/ml to about 1.5 mg/ml.
9. The composition of claim 8 , wherein the recombinant protein is present in an amount of about 1 mg/ml.
10. The composition of claim 9 , wherein the composition further comprises an adjuvant.
11. The composition of claim 10 , wherein the adjuvant is present in an amount of 5-15% by volume.
12. The composition of claim 11 , wherein the adjuvant is present in an amount of about 10% by volume.
13. The composition of claim 12 wherein the adjuvant is a carbomer-based adjuvant.
14. The composition of claim 6 , wherein the carrier is an aqueous solution, and the composition has a pH between about 6.5 and about 7.5.
15. The composition of claim 14 , wherein the pH is between about 6.7 and about 7.2.
16. A composition comprising:
(a) an isolated recombinant protein comprising consecutive amino 29-189 of Rhodococcus equi virulence-associated protein A (VapA) (i) the sequence of which is set forth in SEQ ID NO:1, or (ii) varies from such sequence by not more than five amino acids;
(b) an isolated recombinant protein comprising consecutive amino acids 29-173 of Rhodococcus equi virulence-associated protein (VapC) the sequence of which is set forth in SEQ ID NO:3, or (ii) varies from such sequence by not more than five amino acids; and
(c) a carrier.
17. The composition of claim 16 , wherein the carrier is an aqueous or non-aqueous solution, suspension or emulsion and wherein (a) and (b) are present in an amount which may be the same or different and is between about 0.25 mg/ml and about 2.5 mg/ml.
18. The composition of claim 17 , wherein each of (a) and (b) is present in an amount between about 0.5 mg/ml and about 1.5 mg/ml.
19. The composition of claim 18 , wherein each of (a) and (b) is present in an amount of about 1 mg/ml.
20. The composition of claim 16 , wherein the composition further comprises an adjuvant.
21. The composition of claim 20 , wherein the carrier is an aqueous or non-aqueous solution, suspension or emulsion and wherein the adjuvant is present in an amount of 5-15% by volume.
22. The composition of claim 21 , wherein the adjuvant is present in an amount of about 10% by volume.
23. The composition of claim 20 wherein the adjuvant is a carbomer-based adjuvant.
24. The composition of claim 19 , wherein the carrier is an aqueous solution, and wherein the composition has a pH between about 6.5 and about 7.5.
25. The composition of claim 24 , wherein the pH is between about 6.7 and about 7.2.