APR Nov/Dec 2022 - 65

VENDOR VIEWPOINT
method is an end-point method, meaning one reading is taken at
the beginning of the test, and then one at the very end. It works
by a fluorescence reader amplifying a fluorogenic substrate when
endotoxin is detected. Since this is an alternative method for detection
of bacterial endotoxin, a company must go through additional and
rigorous testing to show equivalency to the compendial BET methods.
The FDA has recognized rFC as an alternative method, and states
firms may use alternative methods and/or procedures if they provide
advantages in terms of accuracy, sensitivity, precision, selectivity, or
adaptability to automation or computerized data reduction, and in
other special circumstances.3
If a company chooses to use the rFC
assay, then alternative method validation should be performed in
accordance with the requirements of USP <85> Bacterial Endotoxins
Test and USP Chapter <1225> Validation of Compendial Procedures.3
Recombinant LAL reagent, or rLAL, is also an alternative test. It utilizes
the same cascade as traditional LAL reagents. This means it contains
all the factors that are responsible for detecting endotoxin that are
found in horseshoe crab blood using a recombinant manufacturing
process.4
This a step forward in the process of using a recombinant
reagent as it contains all clotting factors and uses a traditional kinetic
assay on an absorbance reader. Although it is a step forward in animalfree
testing for BET, it is still considered an alternative method and
companies must go through the same time consuming and rigorous
process as rFC if they choose to use this assay. Since trying to validate
and implement an alternative method for BET is not easy, some firms
may ask what other solutions are currently available to help achieve
a more sustainable and efficient way to test products for endotoxin.
The Sievers Eclipse Bacterial Endotoxins Testing (BET) Platform is one
solution that offers significant advantages in terms of LAL reduction
65
American Pharmaceutical Review | November/December 2022
References
1.
2.
3.
4.
Atlantic States Marine Fisheries Commission. (n.d.). Horseshoe Crab. Retrieved from
http://www.asmfc.org/species/horseshoe-crab
Floyd, B., & DeLancey, L. (2012, March 1). South Carolina Horseshoe Crab Fishery and
Management Program Compliance Report for the Year 2011. Retrieved from https://
www.dnr.sc.gov/marine/mrri/pubs/SCComplianceHSC2011.pdf
U.S. Department of Health and Human Services Food and Drug Administration. (2012,
June). Guidance for Industry: Pyrogen and Endotoxins Testing: Questions and Answers
Associates Of Cape Cod Incorporated. (n.d.). Recombinant LAL Reagent PyroSmart
NextGen™ Instructions For Use.
and compliance. The Eclipse reduces LAL use by up to 90 percent while
remaining compliant with all global pharmaceutical regulations. This
is a substantial improvement compared to traditional LAL methods
such as the 96-well plate and gel clot. For the Sievers Eclipse BET
platform, a novel microplate was designed that facilitates accurate
and rapid dispersion of LAL and samples using centripetal force,
metering chambers, and microfluidic channels. The microfluidic
system enables users to carry out the same biochemistry that is
performed in traditional endotoxin assays but with minimal hands-on
effort, greater consistency, and drastically reduced LAL consumption.
By decreasing horseshoe crab lysate use by up to 90%, the Eclipse
platform reduces the demand on the most sensitive and unmatched
natural endotoxin detection reagent on the planet. The Eclipse
platform uses commercially available, FDA licensed LAL and meets all
requirements of the harmonized global pharmacopoeia, USP <85>,
EP 2.6.14 and JP 4.01. This empowers users to be conscious of today's
needs to protect valuable natural resources while still complying with
the strict analytical and regulatory requirements drug and device
manufacturers must meet.
http://www.asmfc.org/species/horseshoe-crab https://www.dnr.sc.gov/marine/mrri/pubs/SCComplianceHSC2011.pdf
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