Embracing Sustainability with Endosafe® Trillium™

Charles River’s animal-free bacterial endotoxin testing recombinant cascade reagent

Back in May, during our interview with endotoxin expert Nicola Reid, we discussed the evolution of endotoxin tests and considerations for what’s next for the “gold standard” LAL test. Shortly after that chat, we introduced this summer the latest addition to Charles River’s bacterial endotoxin detection portfolio, Endosafe Trillium, an animal-free recombinant cascade Reagent (rCR). To get the inside scoop, we sat down with Courtney Wachtel, Endosafe® Product Manager. Courtney has 10 years of experience in the pharmaceutical industry, and - in her current role - focuses specifically on endotoxin detection assays, with a keen focus in rCR.

Why did Charles River decide to enter the recombinant endotoxin space?

Courtney: In the pursuit of developing an alternative to the horseshoe crab-derived endotoxin test, we've conducted several scientifically sound studies, analyzed real world data, challenged our results, and optimized our reagent formulation. This resulted in the creation of our rCR, Endosafe Trillium, that completes our Endosafe portfolio and aligns with our customers' and the industry's expectations. Some things take time to get right, including strong and precise science. Ensuring the utmost quality, safety, and accuracy in a recombinant cascade reagent matters to us, our customers, and patients. So, for us, it has been worth the wait.

What makes Trillium different than other recombinant products available?

Courtney: Trillium is a 100% animal-free recombinant cascade reagent used for the detection of bacterial endotoxins. It's an optimized kinetic chromogenic recombinant assay that detects natural environmental endotoxin by simulating the natural LAL cascade reaction from the Limulus polyphemus species. Throughout the optimization of our formulation, we not only tested our recombinant cascade reagent alongside reference standard endotoxin and control standard endotoxin, but we also ensured that we were testing natural environmental endotoxin, as well. Our team spent a lot of time understanding the importance of interference patterns and formulation and the impact that those two have on detecting those endotoxins. They took a steady and thorough approach that has allowed us to ensure that our formulation is robust and equivalent to LAL. Lastly, we have ensured that this kinetic chromogenic reagent works with all our current incubating microplate instrumentation and software. This will not only have minimal impact to a lab currently using a chromogenic method, but it also provides a natural amplification of the endotoxin reaction. 

You mentioned that Trillium’s formulation is robust and equivalent to LAL. Would you mind providing more insights on this?

Courtney: Our rCR contains three critical proteins that improve testing accuracy in terms of comparability and robustness when compared to other recombinant endotoxin detection technologies. We firmly believe that there is an evolutionary aspect to the way in which the natural LAL cascade unfolds, and this is why we have included the three main proteins found in the LAL cascade: Factor C, Factor B, and Pro-clotting Enzyme. These three proteins, which we have developed each as a recombinant, allow us to ensure accurate recovery of both NOEs (Naturally Occurring Endotoxin) and NEEs (Natural Environmental Endotoxin).

As we celebrate 20 years of the LAL cartridges, will Trillium be integrated into the flagship cartridge technology?  

Courtney: Yes, we're currently in the process of finalizing the development of our recombinant cartridges and slated to launch within the first quarter of 2024. The same Endosafe software and instrumentation can be utilized in addition to the same testing procedure as currently used for our LAL cartridges, i.e., by simply adding 25 microliters to the four cartridge wells and then running the assay. 

You talk a lot about sustainability, so what does “sustainable endotoxin testing” mean?

Courtney:  We've been hearing that the biggest initiative fueling customer’s labs is animal product reduction or replacement, followed by supply chain protection, and the ability to scale and innovate. Charles River has been focused on sustainable bacterial endotoxin test methods for over 20 years. This year marks the 20th anniversary of our flagship cartridge technology, released in 2003 for R&D usage. Our cartridge technology reduces the LAL reagent utilized per test by 95%. To continue with our sustainability journey, the next step was finalizing the development and releasing a recombinant cascade reagent, allowing us to remove the remaining 5% of LAL in comparison to our standard cartridges. Charles River is committed to providing sustainable options for all our customers’ needs. To underline the importance of our pledge to be sustainable, we have recently reframed the 3Rs and added a fourth R for “Responsibility” which demonstrates our responsibility in making long-term investments that lead the industry, while ensuring patient safety and sustainability.

How can we learn more about Endosafe Trillium?

Courtney: We recently hosted a webinar series in which we explain our Endosafe Trillium and how it works, providing an overview of product test data and results, as well as an explanation of our comprehensive validation protocols and implementation support. Listening to it is an amazing opportunity to understand our latest addition to our BET portfolio. Here is the link to watch it on-demand and have access to other relevant resources.

If you would like to have more information, feel free to reach out by email to [email protected] or [email protected].