Fig. 1: Split luciferase sensors for detecting BoNT activity.
From: Split luciferase-based assay to detect botulinum neurotoxins using hiPSC-derived motor neurons
a Schematic representation of the split luciferase-based biosensor. The split NanoLuc™ luciferase is linked together by a linker containing a SNARE sequence. By cleaving the SNARE part of the sensor, BoNTs separate NanoLuc™ luciferase and prevent it from reconstitution into a functional luciferase. The decrease in luminescence is inversely proportional to the toxin activity. b Schematic drawing of two sensor proteins designed for BoNTs detection. For SNAP-25 (p25)-cleaving BoNT, the sensor N-Nano-p25-C-Nano was prepared. For VAMP1(vp1)-cleaving BoNT, the sensor protein N-Nano-vp1-C-Nano was prepared. c Acellular detection assay of BoNT/A. Sensor proteins (30 nM) were mixed with different concentrations of BoNT/A. The curve was plotted based on the percentage of luminescence of NanoLuc luciferase by using toxin added sample/control sample. Assay was performed in duplicate. d Acellular detection assay of BoNT/B. The process was the same as described for detecting BoNT/A. Assay was performed in duplicate.
