Recombinant Cascade Reagents
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Frequently Asked Questions
1. Will PSNG perform the same as naturally sourced reagents?
Yes, PSNG is a kinetic chromogenic reagent. It contains recombinant Factor C, Factor B and pro-clotting enzyme, thus it continues to rely on the cascade mechanism in order to amplify the signal. It is co-lyophilized with the same chromogenic substrate as other chromogenic reagents. Therefore it is homogeneous with Pyrochrome® and Chromo-LAL. This presents a huge advantage to the end user: they keep the method, only switch out the reagent.
2. What’s the difference between Recombinant Factor C (rFC) and Recombinant Cascade Reagent (rCR)?
Recombinant Factor C reagent contains only recombinant Factor C. This enzyme acts as a biosensor for endotoxin (specifically binds to it). But since the cascade amplifying the signal is missing (no Factor B and pre-clotting enzyme), Factor C reagents are used in combination with fluorescence read out in order to achieve sensitivity levels comparable to the cascade reaction. This means that a fluorescent plate reader has to be used (along with a specific microplate). So this means different method, different readers, different SOPs for rFC reagents. Recombinant cascade reagent (e.g. recombinant TAL (= PyroSmart) or recombinant LAL (= PyroSmart NextGen™) are homogenous to current LAL reagents (Pyrochrome® or Chromo-LAL) in that they contain the full cascade (3-enzyme cascade).
3. Does PSNG use any raw material from the horseshoe crab?
No. PSNG is completely animal free.
4. Will new instrumentation be required to use PSNG?
No. Due to the cascade based feature, you can utilize the same instrument and preparation procedures.
5. How are recombinant reagents produced?
It is a biopharmaceutical production based on the use of expression system as cell line clones, that contain (thanks to genetic engeering) DNA coding Factor C, Factor B and pro-clotting enzymes from Limulus Polyphemus. The cell lines express the final glycoproteins Factor C, Factor B and pro-clotting enzyme which are then collected, purified and formulated into PyroSmart NextGen™.