Recombinant expression of PCV2b ORF2 protein in insect cells
AssigneeIntervet Inc.
InventorsPaulus Jacobus Antonius Sondermeijer, Lisette Sanders, Karin Huberdina Antonia Van Der Heijden-Liefkens
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Abstract
The present invention relates to the field of veterinary vaccines, in particular to porcine vaccines against PCV2 and associated diseases. Specifically the invention relates to the finding that a mutation is required in PCV2b ORF2 protein, to prevent its nuclear accumulation upon expression in insect cells; the mutation introduces a Proline at amino acid position 131. This allows efficient expression in insect cells, easy harvesting, and generates large amounts of virus-like particles. The VLPs are highly effective in vaccines for porcines for reduction of infection by PCV2 or of associated signs of disease.
Claims13
An insect cell comprising a heterologous nucleic acid comprising:
- a. a nucleotide sequence encoding an ORF2 protein from porcine circovirus 2 of genotype 2b (PCV2b), and
- b. a transcription control sequence that is operatively linked to said nucleotide sequence, wherein the nucleotide sequence encodes a mutant PCV2b ORF2 protein having a Proline at amino acid position number 131.
A virus-like particle (VLP) of a mutant PCV2b ORF2 protein, wherein the mutant PCV2b ORF2 protein has a Proline at amino acid position number 131.
A vaccine for porcine animals for reducing infection by PCV2 or reducing associated signs of disease, comprising a mutant PCV2b ORF2 protein in a pharmaceutically acceptable carrier and an adjuvant, wherein the mutant PCV2b ORF2 protein has a Proline at amino acid position number 131.
Dependencies are inferred from the claim text.