V5 peptide – Epitope of Simian Parainfluenza Virus type 5
Simian Parainfluenza Virus type 5
V5 peptide is an epitope peptide of Simian Parainfluenza Virus type 5 (SV5) belonging to the Paramyxovirus family. SV5 causes some common human diseases,such as pneumonia,bronchiolitis,measles and mumps.
V5 peptide
V5 peptide represents amino acids 95-108 of SV5 corresponding to RNA polymerase alpha subunit and non-structural protein V. V5 peptide sequence was chosen because high-affinity antibodies can be produced in many different species. V5 peptide is useful as tag for protein purification and detection such as Western blotting,ELISA assays,immunofluorescence,immunoprecipitation assays…
Two canine isolates of simian virus 5 (SV5),termed CPI+ and CPI-,were examined for their ability to react with a bank of monoclonal antibodies (MAbs) that had been previously raised against a human isolate of SV5. CPI- virus was originally isolated from the brain of a gnotobiotic dog infected with CPI+ virus and establishes persistent infections more readily than CPI+ in vitro. Of more than 50 MAbs tested,only one (P-k) reacted with CPI+ but not CPI-,enabling distinction between the two canine isolates. It had been shown previously that MAb P-k reacts with an epitope common to both the P and V proteins. In order to characterize further the epitope binding site of this MAb the P/V genes of CPI+ and CPI- were sequenced. There were four nucleotide differences between CPI+ and CPI-,three of which resulted in predicted amino acid substitutions. Synthetic peptides corresponding to regions encompassing these changes were made and radioimmune competition assays were used to identify the epitope binding site of MAb P-k. Sequence comparison of the P/V gene of CPI+ with the published sequence of a monkey isolate of SV5 (W3) revealed 14 nucleotide differences with five amino acid substitutions. The only amino acid substitution observed between CPI+,CPI- and W3 which altered the predicted secondary structures of the P and V proteins was a leucine to proline change that induced a predicted beta-turn and resulted in the loss of binding of MAb P-k.
2- Huang L. et al. Virus Res. 161(2):115-123 (2011)
Porcine circovirus type 2 (PCV2) is a major causal agent of post-weaning multisystemic wasting syndrome in piglets. To investigate the feasibility of PCV2 expressing an exogenous epitope,a 14-amino-acid V5 epitope derived from simian parainfluenza virus type 5,was inserted into the C terminus of the capsid protein. Recombinant PCV2 expressing the V5 epitope,recPCV2/CL-V5,was rescued by transfecting an infectious clone into PK-15 cells and was characterised by an immunoperoxidase monolayer assay (IPMA),a serum neutralisation assay (SNA),a capture enzyme-linked immunosorbent assay (ELISA) and immunoelectron microscopy. The V5 epitope was detected in the recombinant marker virus by IPMA and capture ELISA. Furthermore,there was no detectable difference in the antigenicity of the recombinant marker virus compared with the parental virus by IPMA and SNA using PCV2-positive serum and the neutralising monoclonal antibody 1D2. However,recPCV2/CL-V5 marker virus could be differentiated from the parental virus by PCR,IPMA and capture ELISA. The recombinant marker virus was stable on multiplication through 10 passages in PK-15 cells,with a maximum titre of 10(6.25) 50% tissue culture infective dose (TCID(50))/ml. BALB/c mice were inoculated with the recombinant or parental virus via the intranasal and intraperitoneal routes. The parental and recombinant viruses both could replicate in mice,cause microscopic pathological changes,and induce mice to generate anti-PCV2 antibodies. Furthermore,the recombinant marker virus could also induce anti-V5 epitope tag antibodies. These results indicated that V5 epitope could be displayed on the surface of the capsid protein by inserting its gene just before stop codon of open reading frame 2. More importantly,insertion of the V5 epitope did not seem to interfere with biological characterisation of the recPCV2/CL-V5 marker virus.
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