email: [email protected] vaccines and adjuvants url: bam hi(1513) sal i(1527) not i(1520) hn aﬃnity...

Vaccines and Adjuvants Nature Technology Corporation 4701 Innovation Drive Lincoln, NE 68521 Phone: (402) 472-6530 Email: [email protected] URL: www.natx.com Thermostable Protein Expression: Antigen/Adjuvant 1 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 - Influenza Antigen Nde I(1508) pfMBP pVEXK HN-MBP-MCS-K6EE 5811 bp Bam HI(1513) Sal I(1527) Not I(1520) HN affinity tag lac IQ pVEX3 seq Inducible Taq promoter Kan R Terminator K6EE Eco RI(1566) pBR322 origin Optimized mRNA Leader Xba I(247) Enterokinase Gel : NTC developed TLR5 receptor agonist TST-Flagellin. lane 1 = uninduced total protein lane 2 = induced total protein lane 3 = uninduced soluble protein, lane 4 = induced soluble protein lane 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-flagellin). 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 Effective 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 a Absorbance of 1/62,500 dilution ºP<.05 by two sided Student t-test

Upload: others

Post on 07-May-2020

2 views

Category:

Documents

0 download

Report

Download

Embed Size (px):

TRANSCRIPT

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