Marker system, in particular for baculovirus-expressed subunit antigens
The present invention belongs to the field of compliance markers and marker vaccines which allow for the differentiation between infected and vaccinated individuals. In particular, it relates to a method of determining whether an individual has received an immunogenic composition comprising a recombinant protein produced by a baculovirus expression system in cultured insect cells.
1. A method for determining whether an animal has received a marker vaccine including a recombinant protein produced by an expression system in cultured insect cells, comprising:
detecting, in a biological sample derived from the animal, for a marker indicating whether the animal has been exposed to an antigen from an RNA virus capable of infecting insect cells.
2. The method of claim 1 , wherein the RNA virus is a rhabdovirus.
3. The method of claim 2 , wherein the marker is an antibody specific for a rhabdovirus antigen.
4. The method of claim 2 , wherein the antigen is a G protein.
5. The method of claim 2 , wherein the antigen is an N protein.
6. The method of claim 1 , further comprising contacting the marker with a capture reagent under specific hybridization conditions, wherein the capture reagent binds the marker under the specific hybridization conditions to form a marker-capture reagent complex.
7. The method of claim 6 , further comprising contacting the marker-capture reagent complex with a detectable agent that specifically binds to the marker-capture reagent complex.
8. The method of claim 7 , wherein the detectable agent is a labelled secondary antibody.
9. The method of claim 6 , wherein the capture reagent is a rhabdovirus.
10. The method of claim 9 , wherein the rhabdovirus is inactivated.
11. The method of claim 6 , wherein the capture reagent is a baculovirus-expressed protein.
12. The method of claim 6 , wherein the capture reagent is (i) a polypeptide comprising a sequence having at least 95% sequence identity with any one of SEQ ID NOs: 1 to 8, (ii) a synthetic polypeptide consisting of 5 to 11 consecutive amino residues from SEQ ID NO:1, or (iii) a synthetic polypeptide consisting of 5 to 11 consecutive amino residues from SEQ ID NO:7.
13. The method of claim 1 , wherein the animal is a pig or a chicken.
14. The method of claim 1 , wherein the biological sample comprises whole blood, blood plasma, serum, urine, oral fluids, or any combinations thereof.
15. The method of claim 1 , wherein the RNA virus is capable of infecting a Spodoptera Frugiperda cell line.
16. The method of claim 1 , wherein the marker vaccine further comprises the antigen, and wherein the expression system is a baculovirus expression system.
17. The method of claim 16 , wherein the recombinant protein is a PCV2 ORF2 protein having at least 95% sequence identity with SEQ ID NO:23.
18. The method of claim 16 , wherein the recombinant protein is a hemagglutinin of an avian influenza H5N1 virus and has at least 95% sequence identity with SEQ ID NO:24.
19. The method of claim 1 , wherein the marker is (i) the antigen, (ii) an antibody specific for the antigen, and/or (iii) a nucleic acid of the RNA virus.
20. The method of claim 1 , wherein the marker vaccine is a positive marker vaccine, and wherein detecting the marker in the biological sample indicates that the animal has received the marker vaccine.