a brighter day with improved luciferins adapted to ... · a brighter day with improved luciferins...
TRANSCRIPT
A brighter day with improved luciferinsadaptedtoOplophorus-derivedluciferases
Improved bioluminescent signals (using a NanoLuc™/nanoKAZ-based luciferase) obtained with our proprietary luciferins incomparisonwith the current “glowing” standard furimazineorthepreviouslyundisclosed“flashy”hydroxyfurimazine.
Contact [email protected]
TechID 201625Inventor YvesJaninPatent ProcessingPartnering Out-Licensing
Furimazine
Ø New original robust synthetic pathwayØ More than 100 synthesized luciferinsØ Up to x2.5 increased intensityØ Extended stable signal up to 2 hoursØ In vitro& in vivo HTSØ 2D & 3D imaging in cellulo assaysØ 3D imaging in vivo assay
Time(minutes)
RLU
500000
1000000
1500000
2000000
2500000
3000000
020 40 60 80 100 120
Furimazine
Hydroxyfurimazine
A
B
Figure 1 - Bioluminescence emission over two hours offurimazine and hydroxyfurimazine as well as our improvedproprietary luciferins (A) and (B).
Light2Ourproprietary
improvedluciferins
Oplophorus-derivedluciferase
Ourproprietary improvedluciferins (A,B,C,D…)
T-cell
Furim
azine
JurkatHEK
NanoKAZ-IL7 NanoKAZ-IL7 NanoKAZ-IL7
(C)
NanoKAZ-IL7 NanoKAZ-IL7 NanoKAZ-IL7
2minafterinjection of(D)
30minafterinjection of(D)
Figure 2 - Expression and location of proteinchimeras fused to the nanoKAZ in transientlytransfected HEK and Jurkat cell lines, and primaryhuman T-cells. The cytokine receptor IL7Ra:nanoKAZis located at the membrane. Images were acquiredfor one second from live cells using our improvedproprietary luciferin (C) (top line) and furimazineshowing the subcellular resolution in cell cultureconditions and the gain of light emisionwith C.
Figure 3 - Mouse infected with Ross River virus expressing NanoLuc (day 6pi). Intense bioluminescent signal in muscle tissues indicating high viral replication (our improved proprietary luciferin (D) is IP injected, 30s of exposition, medium binning).
Substrate lmax (nm) Imax Half-life
Furimazine 460 1 1
Hydroxyfurimazine - 3.06 <0.05
A - 0.84 2.29
B - 2.30 0.79
C - 2.70 0.36
D - 2.07 0.30
Table 1 - Bioluminescence properties of our proprietary improved luciferins A, B, C and D normalized using Furimazine as reference (used in combination with Oplophorus-derived luciferase, crustacean of the family Oplophoridae).