BPB Reports

Paper Details

BPB Reports
Vol. 5 No. 5 p.105-114 2022
Regular Article
Advanced Recombinant Cascade Reagent PyroSmart NextGen® for Bacterial Endotoxins Test as Described in the Pharmacopeias
  • Ingrid Stevens (R&D Department, Associates of Cape Cod, Inc., USA / istevens@acciusa.com)
  • Norihiko Ogura (LAL Research, Central Research Laboratory, Seikagaku Corporation / norihiko.ogura@seikagaku.co.jp)
Ingrid Stevens 1) , Norihiko Ogura 2) , Madeline Kelley 1) , Robert L. D’Ordine 1) , Hikaru Mizumura 2) , Toshio Oda 2) , Junko Akiyoshi 1) , Edwin G. Jahngen 1)
1) R&D Department, Associates of Cape Cod, Inc., USA , 2) LAL Research, Central Research Laboratory, Seikagaku Corporation
Received: August 26, 2022;   Accepted: September 22, 2022;   Released: October 28, 2022
Keywords: bacterial endotoxins test, recombinant cascade reagent, lysate reagent


The bacterial endotoxins test (BET) is a method of detection and quantification of bacterial endotoxin in injectable drugs and medical devices using amoebocyte lysate reagents sourced from the horseshoe crab (Limulus polyphemus or Tachypleus tridentatus). Three manufacturers have launched three different types of recombinant reagents, and recently the United States, European and Japanese Pharmacopeias have been evaluating the equivalency of these reagents to natural amoebocyte lysate reagents. Several studies suggested that the recombinant reagents are very similar to amoebocyte lysate reagents, however there is potential for improvement. Previous reports indicate that recombinant reagents have two issues: extremely low endotoxin potency determination for Helicobacter pylori GU2 and high levels of interference from Heparin Calcium impacting endotoxin recovery. A new recombinant cascade reagent (rCR), PyroSmart NextGen®, recently introduced to the market has been developed to solve these issues. PyroSmart NextGen® has demonstrated higher reactivity to H. pylori GU2 and a lower level of interference from Heparin Calcium than other existing recombinant reagents. Additionally, the analytical capability and suitability of PyroSmart NextGen® has been demonstrated when applied to the BET as described in the US, European and Japanese pharmacopeias. PyroSmart NextGen® has also shown comparability to amoebocyte lysate reagents by demonstrating its ability to detect autochthonous endotoxin in water and in Escherichia coli culture. Overall, this study has verified that the rCR, PyroSmart NextGen® is a suitable alternative to amoebocyte lysate reagents.