Dual expression system for assembling phage lambda display particle (LDP) vaccine to porcine Circovirus 2 (PCV2).
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.
Hayes
Sidney
S
Department of Microbiology and Immunology, College of Medicine, University of Saskatchewan, 107 Wiggins Road, Saskatoon, SK S7N 5E5 Canada. sidney.hayes@usask.ca
Gamage
Lakshman N A
LN
Hayes
Connie
C
D000595:Amino Acid Sequence; D000818:Animals; D057134:Antibodies, Neutralizing / Q000097:blood; D000914:Antibodies, Viral / Q000097:blood; D000917:Antibody Formation; D010582:Bacteriophage lambda / Q000276:immunology; D001483:Base Sequence; D036022:Capsid Proteins / Q000276:immunology*; D018173:Circoviridae Infections / Q000276:immunology / Q000517:prevention & control*; D017925:Circovirus / Q000276:immunology*; D000939:Epitopes / Q000276:immunology; D005856:Germ-Free Life; D007111:Immunity, Cellular; D008969:Molecular Sequence Data; D019151:Peptide Library; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000276:immunology; D013552:Swine; D014765:Viral Vaccines / Q000276:immunology*
D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D036022:Capsid Proteins; C041745:D protein, Enterobacteria phage lambda; D000939:Epitopes; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D014765:Viral Vaccines