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PHARMALEADS substrates for BoNT quantification
Thanks to their high sensitivity, PHARMALEADS’ enzymatic assays for Botulinum neurotoxins (BoNT) A and B are becoming the reference for BoNT detection and quantification.
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The assays rely on original and patented fluorigenic peptide substrates using our FLUOFAST® technology.
They provide the BoNT research community and industry with a novel, powerful, non-animal based method for BoNT quantification.The fluorogenic couple (Pya-fluorophore / Nop-repressor) is introduced in a synthetic substrate, based on the peptide sequence of either SNAP-25 or VAMP1, the SNAREs naturally cleaved by BoNT/A and BoNT/B respectively.
The fluorogenic Pyrenylalanin (Pya) and repressor Para-nitro-phenylalanin (Nop) moieties flank the specific peptide bond naturally cleaved by the toxins. Combined with the low base fluorescence of the quenched substrate, the high fluorescence of Pya provides an intense signal, allowing a high sensitivity of the assay (below 15 LD50 in optimized conditions).
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A highly specific, sensitive and versatile technology :
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PHARMALEADS substrates1
PL50 [(Ac-156-203) SNAP-25](Nop197, Pya200, Nle202) and
PL51 [(Ac-156-203) SNAP-25](Nle163, Nle167, Nle182, Nop197, Pya200, Nle202) are specific to BoNT/A while
PL150 [(Ac-60-94) VAMP](Pya74,Nop77), is specific to BoNT/B.
There is no cross reactivity with other BoNT serotypes as well as with other proteases.
Technology applicable to any kind of botulinum toxin A and B, from light chain to any kind of high molecular weight toxin.
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Convenient and user friendly technology:
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This single-step assay can be used for quality control of toxin preparations as well as during production processes.
The assay is adapted to match HTS testing requirements in term of sensitivity and cost.
The fast enzymatic reaction allows a quick detection in 60 minutes and/or high sensitive measurement in up to 300 minutes.
A common plate fluorimeter is needed to perform the assay (λex = 340 nm, λem = 405 nm).
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