Highly protective E2-CSFV vaccine candidate produced in the mammary gland of adenoviral transduced goats.
Classical swine fever virus is the etiological agent of the most economically important highly contagious disease of swine worldwide. E2 is the major envelope glycoprotein present as a homodimer on the outer surface of the virus and represents an important target for the induction of neutralizing immune response against the viral infection. The E2 extracellular domain was expressed in the milk of adenoviral transduced goats at the highest level about 1.2g/L. The recombinant glycoprotein was purified from clarified serum milk by a single metal chelate affinity chromatography step, as a homodimer of approximately 100kDa and purity over 98%. Glycosylation analysis showed the presence of oligomannoside, hybrid and complex type N-glycans, attached to the recombinant E2. The capacity of goat milk-derived E2 antigen to protect pigs from both classical swine fever clinical signs and viral infection was assessed in a vaccination and challenge trial. The immunized pigs became protected after challenge with 10(5) LD(50) of a highly pathogenic CSFV strain. In the context of veterinary vaccines, this expression system has the advantages that the recombinant antigen could be harvested in about 48h after adenoviral transduction with expression levels in the range of g/L. This approach may turn into a scalable expression system for the assessment and production of veterinary vaccines.
Animal Biotechnology Department, Centre for Genetic Engineering and Biotechnology (CIGB), Havana, Cuba. jorge.toledo@cigb.edu.cu
Jorge R
JR
Toledo
Oliberto
O
Sánchez
Raquel
R
Montesino
Omar
O
Farnos
Maria P
MP
Rodríguez
Pastor
P
Alfonso
Nayrobis
N
Oramas
Elsa
E
Rodríguez
Elaine
E
Santana
Ernesto
E
Vega
Llilianne
L
Ganges
Maria T
MT
Frias
José
J
Cremata
Maritza
M
Barrera
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D000818:Animals; D001831:Body Temperature; D002460:Cell Line; D006691:Classical Swine Fever / Q000517:prevention & control*; D006692:Classical Swine Fever Virus / Q000276:immunology*; D019281:Dimerization; D006031:Glycosylation; D006041:Goats; D006639:Histidine; D006801:Humans; D007273:Injections, Intramuscular; D008321:Mammary Glands, Animal / Q000378:metabolism*; D008892:Milk / Q000737:chemistry / Q000276:immunology; D009842:Oligopeptides; D009844:Oligosaccharides / Q000378:metabolism; D013552:Swine; D013997:Time Factors; D014161:Transduction, Genetic; D014759:Viral Envelope Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D014765:Viral Vaccines / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
C471213:His-His-His-His-His-His; D009842:Oligopeptides; D009844:Oligosaccharides; D014759:Viral Envelope Proteins; D014765:Viral Vaccines; D006639:Histidine