tetano
Editor, Senior Moderator
Vaccine. 2014 Aug 12. pii: S0264-410X(14)01074-3. doi: 10.1016/j.vaccine.2014.07.092. [Epub ahead of print]
Characterization and immunogenicity in mice of recombinant influenza haemagglutinins produced in Leishmania tarentolae.
Pion C1, Courtois V2, Husson S3, Bernard MC4, Nicolai MC5, Talaga P6, Trannoy E7, Moste C8, Sodoyer R9, Legastelois I10.
Author information
Abstract
The membrane displayed antigen haemagglutinin (HA) from several influenza strains were expressed in the Leishmania tarentolae system. This non-conventional expression system based on a parasite of lizards, can be readily propagated to high cell density (>108cells/mL) in a simple incubator at 26?C. The genes encoding HA proteins were cloned from six influenza strains, among these being a 2009 A/H1N1 pandemic strain from swine origin, namely A/California/07/09(H1N1). Soluble HA proteins were secreted into the cell culture medium and were easily and successfully purified via a His-Tag domain fused to the proteins. The overall process could be conducted in less than 3 months and resulted in a yield of approximately 1.5-5mg of HA per liter of biofermenter culture after purification. The recombinant HA proteins expressed by L. tarentolae were characterized by dynamic light scattering and were observed to be mostly monomeric. The L. tarentolae recombinant HA proteins were immunogenic in mice at a dose of 10μg when administered twice with an oil-in-water emulsion-based adjuvant. These results suggest that the L. tarentolae expression system may be an alternative to the current egg-based vaccine production.
Copyright ? 2014. Published by Elsevier Ltd.
KEYWORDS:
Haemagglutinin; Influenza; Leishmania tarentolae
PMID:
25131728
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/25131728
Characterization and immunogenicity in mice of recombinant influenza haemagglutinins produced in Leishmania tarentolae.
Pion C1, Courtois V2, Husson S3, Bernard MC4, Nicolai MC5, Talaga P6, Trannoy E7, Moste C8, Sodoyer R9, Legastelois I10.
Author information
Abstract
The membrane displayed antigen haemagglutinin (HA) from several influenza strains were expressed in the Leishmania tarentolae system. This non-conventional expression system based on a parasite of lizards, can be readily propagated to high cell density (>108cells/mL) in a simple incubator at 26?C. The genes encoding HA proteins were cloned from six influenza strains, among these being a 2009 A/H1N1 pandemic strain from swine origin, namely A/California/07/09(H1N1). Soluble HA proteins were secreted into the cell culture medium and were easily and successfully purified via a His-Tag domain fused to the proteins. The overall process could be conducted in less than 3 months and resulted in a yield of approximately 1.5-5mg of HA per liter of biofermenter culture after purification. The recombinant HA proteins expressed by L. tarentolae were characterized by dynamic light scattering and were observed to be mostly monomeric. The L. tarentolae recombinant HA proteins were immunogenic in mice at a dose of 10μg when administered twice with an oil-in-water emulsion-based adjuvant. These results suggest that the L. tarentolae expression system may be an alternative to the current egg-based vaccine production.
Copyright ? 2014. Published by Elsevier Ltd.
KEYWORDS:
Haemagglutinin; Influenza; Leishmania tarentolae
PMID:
25131728
[PubMed - as supplied by publisher]
http://www.ncbi.nlm.nih.gov/pubmed/25131728