a brighter day with improved luciferins adapted to ... · a brighter day with improved luciferins...

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 JessicaRobertLicensingManagerJessica.robert@pasteur.fr

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).