Easy and efficient protocols for working with recombinant vaccinia virus MVA.
Modified vaccinia virus Ankara (MVA) is a highly attenuated and replication-deficient strain of vaccinia virus that is increasingly used as vector for expression of recombinant genes in the research laboratory and in biomedicine for vaccine development. Major benefits of MVA include the clear safety advantage compared to conventional vaccinia viruses, the longstanding experience in the genetic engineering of the virus, and the availability of established procedures for virus production at an industrial scale. MVA vectors can be handled under biosafety level 1 conditions, and a multitude of recombinant MVA vaccines has proven to be immunogenic and protective when delivering various heterologous antigens in animals and humans. In this chapter we provide convenient state-of-the-art protocols for generation, amplification, and purification of recombinant MVA viruses. Importantly, we include methodology for rigid quality control to obtain best possible vector viruses for further investigations including clinical evaluation.
Institute for Infectious Diseases and Zoonoses, University of Munich LMU, Munich, Germany.
Melanie
M
Kremer
Asisa
A
Volz
Joost H C M
JH
Kreijtz
Robert
R
Fux
Michael H
MH
Lehmann
Gerd
G
Sutter
D000818:Animals; D015153:Blotting, Western; D002642:Chick Embryo; D003001:Cloning, Molecular; D003264:Containment of Biohazards; D006224:Cricetinae; D016895:Culture Media, Serum-Free; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D016133:Polymerase Chain Reaction; D011786:Quality Control; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014616:Vaccinia virus / Q000235:genetics* / Q000254:growth & development / Q000302:isolation & purification; D019562:Viral Load; D010948:Viral Plaque Assay; D014776:Virus Cultivation
D016895:Culture Media, Serum-Free; D004279:DNA, Viral; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins