tetano
Editor, Senior Moderator
Vaccine
Volume 31, Issue 42, 1 October 2013, Pages 4736?4743
Cover image
Improving influenza virus backbones by including terminal regions of MDCK-adapted strains on hemagglutinin and neuraminidase gene segments
Ra?l C. Gomila E-mail the corresponding author,
Pirada Suphaphiphat E-mail the corresponding author,
Casey Judge E-mail the corresponding author,
Terika Spencer E-mail the corresponding author,
Annette Ferrari E-mail the corresponding author,
Yingxia Wen E-mail the corresponding author,
Giuseppe Palladino E-mail the corresponding author,
Philip R. Dormitzer E-mail the corresponding author,
Peter W. MasonCorresponding author contact information, E-mail the corresponding author
Novartis Vaccines and Diagnostics, 45 Sidney Street, Cambridge, MA 02139, United States
Highlights
?
Chimeric HA and NA gene segments with terminal regions from MDCK-adapted strains were created.
?
Reassortant viruses created with these chimeric segments produced more HA.
?
Combining HA and NA chimeric segments proved superior to chimeric HA or NA only.
Abstract
Reverse genetics approaches can simplify and accelerate the process of vaccine manufacturing by combining the desired genome segments encoding the surface glycoproteins from influenza strains with genome segments (backbone segments) encoding internal and non-structural proteins from high-growth strains. We have developed three optimized high-growth backbones for use in producing vaccine seed viruses for group A influenza strains. Here we show that we can further enhance the productivity of our three optimized backbones by using chimeric hemagglutinin (HA) and neuraminidase (NA) genome segments containing terminal regions (non-coding regions (NCRs) and coding regions for the signal peptide (SP), transmembrane domain (TMD), and cytoplasmic tail (CT)) from two MDCK-adapted high growth strains (PR8x and Hes) and the sequences encoding the ectodomains of the A/Brisbane/10/2010 (H1N1) HA and NA proteins. Viruses in which both the HA and NA genome segments had the high-growth terminal regions produced higher HA yields than viruses that contained one WT and one chimeric HA or NA genome segment. Studies on our best-performing backbone indicated that the increases in HA yield were also reflected in an increase in HA content in partially purified preparations. Our results show that the use of chimeric HA and NA segments with high-growth backbones is a viable strategy that could improve influenza vaccine manufacturing. Possible mechanisms for the enhancement of HA yield are discussed.
http://www.sciencedirect.com/science/article/pii/S0264410X13011171
Volume 31, Issue 42, 1 October 2013, Pages 4736?4743
Cover image
Improving influenza virus backbones by including terminal regions of MDCK-adapted strains on hemagglutinin and neuraminidase gene segments
Ra?l C. Gomila E-mail the corresponding author,
Pirada Suphaphiphat E-mail the corresponding author,
Casey Judge E-mail the corresponding author,
Terika Spencer E-mail the corresponding author,
Annette Ferrari E-mail the corresponding author,
Yingxia Wen E-mail the corresponding author,
Giuseppe Palladino E-mail the corresponding author,
Philip R. Dormitzer E-mail the corresponding author,
Peter W. MasonCorresponding author contact information, E-mail the corresponding author
Novartis Vaccines and Diagnostics, 45 Sidney Street, Cambridge, MA 02139, United States
Highlights
?
Chimeric HA and NA gene segments with terminal regions from MDCK-adapted strains were created.
?
Reassortant viruses created with these chimeric segments produced more HA.
?
Combining HA and NA chimeric segments proved superior to chimeric HA or NA only.
Abstract
Reverse genetics approaches can simplify and accelerate the process of vaccine manufacturing by combining the desired genome segments encoding the surface glycoproteins from influenza strains with genome segments (backbone segments) encoding internal and non-structural proteins from high-growth strains. We have developed three optimized high-growth backbones for use in producing vaccine seed viruses for group A influenza strains. Here we show that we can further enhance the productivity of our three optimized backbones by using chimeric hemagglutinin (HA) and neuraminidase (NA) genome segments containing terminal regions (non-coding regions (NCRs) and coding regions for the signal peptide (SP), transmembrane domain (TMD), and cytoplasmic tail (CT)) from two MDCK-adapted high growth strains (PR8x and Hes) and the sequences encoding the ectodomains of the A/Brisbane/10/2010 (H1N1) HA and NA proteins. Viruses in which both the HA and NA genome segments had the high-growth terminal regions produced higher HA yields than viruses that contained one WT and one chimeric HA or NA genome segment. Studies on our best-performing backbone indicated that the increases in HA yield were also reflected in an increase in HA content in partially purified preparations. Our results show that the use of chimeric HA and NA segments with high-growth backbones is a viable strategy that could improve influenza vaccine manufacturing. Possible mechanisms for the enhancement of HA yield are discussed.
http://www.sciencedirect.com/science/article/pii/S0264410X13011171