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

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 rapid...

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Autores principales: Pasqua, Salvatore, Niotta, Maria Concetta, Di Martino, Giuseppina, Sottile, Davide, Douradinha, Bruno, Miele, Monica, Timoneri, Francesca, Di Bella, Mariangela, Cuscino, Nicola, Di Bartolo, Chiara, Conaldi, Pier Giulio, D’Apolito, Danilo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8408548/
https://www.ncbi.nlm.nih.gov/pubmed/34514024
http://dx.doi.org/10.1016/j.omtm.2021.05.002
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author Pasqua, Salvatore
Niotta, Maria Concetta
Di Martino, Giuseppina
Sottile, Davide
Douradinha, Bruno
Miele, Monica
Timoneri, Francesca
Di Bella, Mariangela
Cuscino, Nicola
Di Bartolo, Chiara
Conaldi, Pier Giulio
D’Apolito, Danilo
author_facet Pasqua, Salvatore
Niotta, Maria Concetta
Di Martino, Giuseppina
Sottile, Davide
Douradinha, Bruno
Miele, Monica
Timoneri, Francesca
Di Bella, Mariangela
Cuscino, Nicola
Di Bartolo, Chiara
Conaldi, Pier Giulio
D’Apolito, Danilo
author_sort Pasqua, Salvatore
collection PubMed
description 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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spelling pubmed-84085482021-09-10 Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes Pasqua, Salvatore Niotta, Maria Concetta Di Martino, Giuseppina Sottile, Davide Douradinha, Bruno Miele, Monica Timoneri, Francesca Di Bella, Mariangela Cuscino, Nicola Di Bartolo, Chiara Conaldi, Pier Giulio D’Apolito, Danilo Mol Ther Methods Clin Dev Original Article 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. American Society of Gene & Cell Therapy 2021-05-14 /pmc/articles/PMC8408548/ /pubmed/34514024 http://dx.doi.org/10.1016/j.omtm.2021.05.002 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Original Article
Pasqua, Salvatore
Niotta, Maria Concetta
Di Martino, Giuseppina
Sottile, Davide
Douradinha, Bruno
Miele, Monica
Timoneri, Francesca
Di Bella, Mariangela
Cuscino, Nicola
Di Bartolo, Chiara
Conaldi, Pier Giulio
D’Apolito, Danilo
Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes
title Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes
title_full Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes
title_fullStr Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes
title_full_unstemmed Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes
title_short Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes
title_sort complete intra-laboratory validation of a lal assay for bacterial endotoxin determination in ebv-specific cytotoxic t lymphocytes
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8408548/
https://www.ncbi.nlm.nih.gov/pubmed/34514024
http://dx.doi.org/10.1016/j.omtm.2021.05.002
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