4.5 Article

Dual expression system for assembling phage lambda display particle (LDP) vaccine to porcine Circovirus 2 (PCV2)

Journal

VACCINE
Volume 28, Issue 41, Pages 6789-6799

Publisher

ELSEVIER SCI LTD
DOI: 10.1016/j.vaccine.2010.07.047

Keywords

Bacteriophage lambda D-fusion expression system; Lambda surface display particle (LDP); vaccine; Porcine Circovirus 2; PCV ORF2 capsid protein

Funding

  1. NSERC Canada [138296]
  2. Western College of Veterinary Medicine

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The bacteriophage lambda small capsid protein D forms trimers on the phage head. D-fusion polypeptides can be expressed from plasmids in E. coli and remain soluble without aggregation. We report a dual expression system for the display of four immunodominant regions of porcine Circovirus 2 (PCV2) capsid protein (CAP) as D-CAP fusions on lambda display particles (LDP). The LDP-D-CAP preparation proved an effective vaccine in pigs, eliciting both cellular and humoral immune responses and PCV2 neutralizing antibodies. In our dual system wild type D expression was encoded by a heteroimmune infecting phage. The D-fusion protein expression in the infected cells was from an inducible plasmid, enabling the deferral of D-fusion expression until needed. The effective vaccine preparation depended upon the gradient purification of very high concentration, essentially tail-less display particles, not previously described. (C) 2010 Elsevier Ltd. All rights reserved.

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