Evaluation of limulus amebocyte lysate and recombinant endotoxin alternative assays for an assessment of endotoxin detection specificity

Dubczak, John; Reid, Nicola; Tsuchiya, Masaharu

doi:10.1016/j.ejps.2021.105716

Cited by 12 publications

(12 citation statements)

References 4 publications

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“…This buffer contains a high concentration of carboxymethylated curdlan, and Charles River suggests its use to prevent the activation of the Factor G zymogen in LAL. 10,11 We then tested the 3 batches of Matrix 1 at the expected dilutions, performing a prior 1:2 dilution with the BG120 buffer. Treatment with BG120 eliminated the interferences, since acceptability criteria in all 3 lots of Matrix 1 were observed in all tests performed at MVD/2 and MVD, showing reproducible and robust data and the validity of the tests (Table 5).…”

Section: Validation Of the Analytical Methods For Both Matricesmentioning

confidence: 99%

Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes

Pasqua

Niotta

Martino

et al. 2021

Molecular Therapy - Methods & Clinical Development

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Endotoxin content is a critical factor that affects the safety of biological pharmaceutical products. International pharmacopoeias describe several reference methods to determine endotoxin levels in advanced therapy medicinal product (ATMP) preparations. Administration of ATMPs must be done as rapidly as possible to ensure complete viability and potency of the cellular product. To evaluate the endotoxin content in the shortest time possible, we chose to validate an alternative method based on the use of the Charles River Portable Testing System (PTS) and FDA-approved cartridges, compliant with the requirements of the European Pharmacopoeia and providing results in <20 min. Here, we describe a unique and complete validation approach for instrument, personnel, and analytical method for assessment of endotoxins in ATMP matrices. The PTS system provides high sensitivity and fast quantitative results and uses less raw material and accessories compared with compendial methods. It is also less time consuming and less prone to operator variability. Our validation approach is suitable for a validated laboratory with trained personnel capable of conducting the ATMP release tests, and with very low intra-laboratory variability, and meets the criteria required for an alternative approach to endotoxin detection for in-process and product-release testing of ATMPs.

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Section: Validation Of the Analytical Methods For Both Matricesmentioning

confidence: 99%

Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes

Pasqua

Niotta

Martino

et al. 2021

Molecular Therapy - Methods & Clinical Development

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“…[16][17][18] Several participants in the roundtable discussion noted published studies that consistently show comparability of rFC-based and LAL-based BET methods for quantifying endotoxin in 'autochthonous' samples, and/or demonstrate advantages to the use of rFC-based methods. [19][20][21][22][23][24][25] In addition, it was noted that a recent study concluding that rFC is not ready for use 26 was published by an LAL supplier. Multiple participants expressed concern that LAL suppliers have previously published similar claims in journals known to deviate from best editorial and publication practices.…”

Section: Scientific Considerations For the Adoption Of Rfc-based Bet ...mentioning

confidence: 99%

“…A recent study published by an LAL supplier 26 was particularly controversial among participants. Several participants questioned the lack of consideration of the variability of LAL-based BET methods in the study design.…”

Section: Recombinant Bacterial Endotoxins Test (Rbet) Roundtable Disc...mentioning

confidence: 99%

Barriers to the Use of Recombinant Bacterial Endotoxins Test Methods in Parenteral Drug, Vaccine and Device Safety Testing

Baker,

Ponder,

Oberdorfer

et al. 2023

Altern Lab Anim

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The Bacterial Endotoxins Test (BET) is a critical safety test that is used to detect bacterial endotoxins, which are the major contributor to fever-inducing contamination risks known as pyrogens. All parenteral therapies, including every lot of injected drugs, vaccines, medical devices, must be tested for pyrogens to ensure patient safety. Bacterial endotoxins test methods were developed as a highly sensitive detection method for bacterial endotoxins, after the discovery of a clotting cascade in horseshoe crab blood. However, horseshoe crab species are limited to some inshore coastal habitats along the Atlantic coast of the USA and others throughout Asia. Fully functional horseshoe crab clotting factors can be manufactured via recombinant protein production, and several BET methods featuring recombinant horseshoe crab proteins have now been developed for commercial use. Recombinant Bacterial Endotoxins Test (rBET) methods based on the use of recombinant Factor C (rFC) were established in the European Pharmacopoeia — however, these methods have not yet been granted compendial status in the United States Pharmacopoeia (USP). In order to facilitate dialogue between stakeholders, the Physicians Committee for Responsible Medicine hosted two virtual roundtable discussions on the perceived barriers to the use of rBET methods for US FDA requirements. Stakeholders agreed that multiple rFC-based methods have been demonstrated to have suitable analytical performance, as described in ICH Q2 on the Validation of Analytical Procedures and USP <1225> on the Validation of Compendial Procedures. United States Pharmacopoeia compendial inclusion of the rFC-based and other rBET methods was favoured, in order to reduce the additional burdens created by a lack of global harmonisation on BET testing requirements.

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“…However, before the blending process can begin, the intermediate endotoxin and potency should be tested. The most commonly recognised tests in industry are Limulus Amebocyte Lysate (LAL) for endotoxin evaluation [357] and plaque formation assays, endpoint dilution assays (tissue culture infective dose TCID 50 ) virus neutralisation assays or quantitative polymerase chain reaction (PCR) assays for potency [358]. The purpose of this evaluation before the blending process is that intermediates with high potencies or endotoxin can be blended with intermediates with low potencies or endotoxin to meet the specification set out in the license.…”

Section: Formulation Considerationsmentioning

confidence: 99%

Nanovaccine Delivery Approaches and Advanced Delivery Systems for the Prevention of Viral Infections: From Development to Clinical Application

et al. 2021

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Viral infections causing pandemics and chronic diseases are the main culprits implicated in devastating global clinical and socioeconomic impacts, as clearly manifested during the current COVID-19 pandemic. Immunoprophylaxis via mass immunisation with vaccines has been shown to be an efficient strategy to control such viral infections, with the successful and recently accelerated development of different types of vaccines, thanks to the advanced biotechnological techniques involved in the upstream and downstream processing of these products. However, there is still much work to be done for the improvement of efficacy and safety when it comes to the choice of delivery systems, formulations, dosage form and route of administration, which are not only crucial for immunisation effectiveness, but also for vaccine stability, dose frequency, patient convenience and logistics for mass immunisation. In this review, we discuss the main vaccine delivery systems and associated challenges, as well as the recent success in developing nanomaterials-based and advanced delivery systems to tackle these challenges. Manufacturing and regulatory requirements for the development of these systems for successful clinical and marketing authorisation were also considered. Here, we comprehensively review nanovaccines from development to clinical application, which will be relevant to vaccine developers, regulators, and clinicians.

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Evaluation of limulus amebocyte lysate and recombinant endotoxin alternative assays for an assessment of endotoxin detection specificity

Cited by 12 publications

References 4 publications

Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes

Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes

Barriers to the Use of Recombinant Bacterial Endotoxins Test Methods in Parenteral Drug, Vaccine and Device Safety Testing

Nanovaccine Delivery Approaches and Advanced Delivery Systems for the Prevention of Viral Infections: From Development to Clinical Application

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