[ACC] Product Overview

KINETIC CHROMOGENIC METHOD RECOMBINANT REAGENT

PyroSmart NextGen ® is a sustainable recombinant Cascade Reagent (rCR) that delivers the same reliable results as your conventional LAL reagent and offers these additional advantages: • No Animal Content – Horseshoe Crab Blood Free • Same Cascade • No Cross Reactivity With 1,3-b-D-glucans • Same Instrument • Same Preparation Steps • Meets Your Sustainability Objectives Keep Your Method. Make An Impact.

ACC’s PyroSmart NextGen ® uses the same cascade as traditional LAL reagents by manufacturing the Factors responsible for the cascade using recombinant processes. As a result, our new recombinant reagent’s mechanism of action will deliver results consistent with traditional LAL reagents. It offers the added advantage of elim- inating 1,3-b-D-glucans cross reactivity from the LAL cascade, since there is no Factor G in the final reagent. ACC developed Pyrosmart NextGen ® to provide a sustainable alternative to traditional naturally sourced LAL reagents, while maintaining your lab procedures, methods, instrumentation and most importantly your results.

The Importance of Mechanism of Action Recombinant Cascade Reagent (rCR)

Recombinant Cascade Reagent (rCR)

Recombinant Factor C Reagent (rFC)

Traditional LAL Reagent

Endotoxin

rFC

Glucan

Glucan No Factor G

rFB

Fluorogenic Substrate

Product Fluorescence

r Proclotting Enzyme

r Clotting Enzyme

Proclotting Enzyme

Clotting Enzyme

Coagulogen Turbidimetric

Coagulin Turbidity Product Yellow Color

Chromogenic

Product Yellow Color

Chromogenic Substrate

Chromogenic

Chromogenic Substrate

Traditional LAL reagent In the presence of endotoxin, Factor C becomes an activated moiety which in turn activates Factor B and Proclotting Enzyme; ultimately result- ing in the proteolytic cleavage of a substrate (either coagulogen in gel clot and turbidimetric assays or a colorless chromogenic substrate in chromogenic assays). The cascade mechanism thus amplifies the response of Factor C and leads to an excep- tional sensitivity for this biological assay, with kinetic output being preferable. In the pres- ence of 1,3- b - D -glucans, Factor G becomes an activated moiety which also activates Proclotting Enzyme and thus resulting in the same signal as that triggered by endotoxins through Factor C. This has been often observed as glucan-derived enhancement or false positive results.

Recombinant Cascade Reagent (rCR) As with naturally sourced LAL reagents, in the presence of endotoxin, recombinant Factor C becomes an activated moiety which in turn activates recombinant Factor B and recombinant Proclotting Enzyme; ultimately resulting in the proteolytic cleavage of a colorless chromogenic substrate formulated with PyroSmart NextGen ® . By relying on the same cascade mechanism, the response of recombinant Factor C is amplified the same way as by LAL reagents and thus the same sensitivity is achieved using this kinetic assay. Due to absence of Factor G, PyroSmart NextGen ® will not react with any 1,3- b - D -glucans and therefore will prevent glucan-derived enhancement and false positive results.

Recombinant Factor C (rFC) – Competition

Launched almost two decades ago, rFC reagents rely only on a recombinant form of Factor C. Due to the absence of the cascade as the amplification mech- anism, rFC reagents are paired with a fluorescence method instead. However, this constitutes a dif- ferent measured entity, different instrumentation, and different preparation steps with a limited output (endpoint assay only). Therefore the uptake and implementation of this method has been rather limited.