Brendan Tindall - bioMérieux
The global pharmaceutical market experienced significant growth in 2020. Accelerated by the ongoing SARS-CoV-2 pandemic, eight vaccines were approved globally with roughly 5.95 billion doses administered and billions more to come.1
The growth of current, emerging and re-emerging infectious diseases has led to the development and approval of new innovative therapeutic approaches, defined as Advanced Therapy Medicinal Products (ATMPs). As ATMP production rises, so does the demand for sustainable and secure Bacterial Endotoxin Testing (BET) in QC labs.
ATMP manufacturers are also being faced with changes in QC-testing needs due to the time-critical, patient-centric manufacturing process (for example, CAR-T cell therapies), alongside demand for more flexible and low-throughput technologies. This is especially true for BET. More importantly, there are also realistic doubts that the currently-used, animal-based BET techniques (namely Limulus Amebocyte Lysate (LAL)) will be able to meet this demand.2
Luckily, the discovery of Recombinant Factor C (rFC) twenty years ago has brought significant progress to the BET techniques of today. This recombinant version can replace the endotoxin-sensitive enzyme, known as LAL, that has historically been extracted from the Horseshoe Crab’s hemolymph. rFC, when compared with LAL-based assays, offers some major and important paradigm improvements: (1) a comprehensive scientific characterization (i.e., fully-characterized reagents); (2) improved specificity through a lack of known or potentially unknown detection pathways for microbe-associated molecular patterns (MAMPs) (i.e., factor G as biosensor for glucans); (3) and a full compliance with the pharmaceutical industry 3R (Reduce/ Refine/Replace) principles, hence sustainability.
In 2018, bioMérieux brought to the market the ENDOZYME II GO assay, a ready-to-use version of the ENDOZYME II assay. Our flagship ENDOZYME II GO assay includes the GOPLATE, a 96 well pre-coated microtiter plate with all the required Reference Standard Endotoxin (RSE)/Control Standard Endotoxin (CSE) standards needed, including a 5-point standard curve from 0.005 – 50 EU/ml and positive product controls with a concentration of 0.5 EU/ml (PPC). This GOPLATE eliminates the need for manual preparation of standard dilutions and PPCs. It also contributes to an average reduction of up to 50% in handling time compared to conventional, LAL-based microplate BET assays. A peer-reviewed industry study shows that the ENDOZYME II GO assay was able to significantly reduce the rate of invalid results when compared with LAL microplate or LAL Cartridge technologies3 (Figure 1).
Flexibility and scalability with minimal waste has been difficult to achieve with regular LAL microplate endotoxin testing solutions. As LAL is not stable in liquid form, it needs to be stored lyophilized with the entire vial to be reconstituted prior usage. In addition, the 96-well microplate allows for only one assay whether you test three samples or 20, thereby potentially wasting unused plates. So, what is the solution?
The new ENDOZYME II GO STRIPS assay offers all the benefits of ENDOZYME II GO, but with the additional flexibility, scalability and waste reduction capability of a modular, strip-based system.
ENDOZYME II GO STRIPS comes with two types of CSE pre-coated modular plates: CURVE STRIPS and SAMPLE STRIPS. The CURVE STRIPS (Figure 2) consist of a 5-point standard curve, blank wells and sufficient wells to analyze one sample including their corresponding PPCs, all in duplicate.
The SAMPLE STRIPS consists of sufficient wells to analyze four samples with their corresponding PPCs in duplicate. This allows you to add as many STRIPS for the samples you have. (Figure 3).
With the liquid rFC reagents provided in each kit, you only use the volume you need for each assay, resulting in a reduction of wasted reagents.
The maximum flexibility of the ENDOZYME II GO STRIPS load-and-go, clickable system ensures complete flexibility for the number of samples you want to test with every assay. For instance, if you have one sample, you only need a single CURVE STRIP; if you have five samples you need to combine one CURVE STRIP and one SAMPLE STRIP. If you have more samples, it`s as easy as clicking another SAMPLE STRIP in place.
ENDOZYME II GO STRIPS also allows for rapid scalability with one reader. Whether you are running one to five samples or 60 samples a day. All can be done with ease with the new ENDOZYME II GO STRIPS assay.
References
- https://ourworldindata.org/covid-vaccinations
- https://www.pda.org/pda-letter-portal/home/full-article/covid-19-and-the-need-for-an- lal-alternative
- Marius M, Vacher F, Bonnevay T. Comparison of bacterial endotoxin testing methods in purified pharmaceutical water matrices. Biologicals 67, 49–55 (2020).
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