A Demonstration of the Validation Process for Alternative Endotoxin Testing Methods Using PyroSmart NextGen<sup>®</sup> Recombinant Cascade Reagent

The Bacterial Endotoxins Test (BET) uses Limulus amoebocyte lysate (LAL) reagents derived from the blood of horseshoe crabs for detection and quantification endotoxins Gram-negative bacteria in parenteral drug products medical devices. Two types recombinant using genes cloned crab genome have become available several suppliers. One is a Factor C reagent (rFC), containing only C, second Cascade Reagent (rCR) factor B proclotting enzyme. Implementation these BET requires validation that demonstrates results equal to or better than those determined by LAL reagents. Previous studies shown PyroSmart NextGen® rCR meets analytical performance requirements both plate tube reader testing methods provides equivalent when samples autochthonous endotoxin. This study directly compares drug, which critical step end-user implementation alternative methods. It first published demonstration an approach includes specific sample, data indicates more precise compared Relative recovery, linear regression, Bland-Altman plot analyses also illustrate are naturally sourced useful method quantifying bacterial drugs.