Vaccines and Adjuvants
Nature Technology Corporation4701 Innovation Drive
Lincoln, NE 68521Phone: (402) 472-6530Email: [email protected]
URL: www.natx.com
Thermostable Protein Expression: Antigen/Adjuvant1
Advantages:
• Thermostable TAG (TST) (maltodextrin binding protein, MBP)
• N-terminal HN affinity tag
• C-terminal K6EE stability/solubility tag
• Inducible Tac promoter
• Optimized mRNA leader
• LacIQ repressor
• Thermostable vaccine components - Flagellin Adjuvant - In�uenza Antigen
Nde I(1508)
pfMBP
pVEXK HN-MBP-MCS-K6EE5811 bp
Bam HI(1513)
Sal I(1527) Not I(1520)
HN affinity tag
lac IQ
pVEX3 seq
Inducible Taq promoter
Kan R
TerminatorK6EE
Eco RI(1566)
pBR322 origin
Optimized mRNA LeaderXba I(247)
Enterokinase
Gel : NTC developed TLR5 receptor agonist TST-Flagellin. lane 1 = uninduced total proteinlane 2 = induced total proteinlane 3 = uninduced soluble protein,lane 4 = induced soluble proteinlane 5 = induced total protein (large scale fermentation harvest)lane 6 = induced soluble protein (large scale fermentation harvest)lane 7 = induced soluble protein after 75ºC 40 min heat treatment (large scale fermentation harvest)M= Marker
Flagellin, a TLR5 agonist, was fused to TST MBP from Pyrococcus furiosus (TST-�agellin).
1) Luke, J., Carnes, A.E., Sun, P., Hodgson, C.P., Waugh, D.S., and Williams, J.A (2011), Thermostable Tag (TST) protein expression system: engineering thermotolerant recombinant proteins and vaccines, J. Biotechnol., 151:242-50.
Thermostable Vaccine Components (larger quantities available by quotation):
HN-flagellin 1mg protein $335
HN-TST-flagellin 1mg protein $335
HN-TST-HA2 1mg protein $335
Pyrococcus furiosus MBP-HA2 is an E�ective Immunogen
Immunogen 56 day antI-HA2 IgG (total) (Abs)a
pfMBP-HA2
HA plasmid
Control
0.615 +/- 0.245º
0.190 +/- 0.138º
0.056 +/- 0.002
aAbsorbance of 1/62,500 dilutionºP<.05 by two sided Student t-test
