An improved method for the recovery of recombinant paramyxovirus vaccine candidates suitable for use in human clinical trials.
We describe a method for the generation of clinical grade, live-attenuated vaccines in Vero cells entirely from cDNA plasmids. The entire electroporation procedure can be completed in less than 15 minutes and this is a significant improvement over previous lipid or electroporation based transfection techniques that also involve a heat-shock step. Importantly, the virus preparations can be generated with a minimal use of animal product derived materials, an important consideration for a vaccine candidate that is to be tested in humans. Since it is likely that all live-attenuated parainfluenza virus and pneumovirus vaccines in the future will be generated using reverse genetics, this simplified method provides guidance on how this can be achieved.
Surman
Sonja R
SR
Laboratory of Infectious Diseases, Respiratory Viruses Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD, USA.
Collins
Peter L
PL
Murphy
Brian R
BR
Skiadopoulos
Mario H
MH
D000818:Animals; D002522:Chlorocebus aethiops; D002986:Clinical Trials as Topic; D018076:DNA, Complementary; D018274:Electroporation; D006801:Humans; D007700:Kinetics; D010222:Parainfluenza Virus 1, Human / Q000235:genetics* / Q000276:immunology; D010223:Parainfluenza Virus 2, Human / Q000235:genetics* / Q000276:immunology; D010224:Parainfluenza Virus 3, Human / Q000235:genetics* / Q000276:immunology; D018098:Paramyxovirinae / Q000235:genetics / Q000276:immunology*; D010957:Plasmids; D011995:Recombination, Genetic; D014613:Vaccines, Attenuated / Q000235:genetics / Q000276:immunology; D014709:Vero Cells; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology*
D018076:DNA, Complementary; D014613:Vaccines, Attenuated; D014765:Viral Vaccines