Recombinant Factor C as an In Vitro Assay for the Residual Pathogenicity Evaluation of Veterinary Autogenous Vaccines

Background: Veterinary autogenous vaccines, similar to all injectable pharmaceutical products, must be tested to assess endotoxin concentrations. The Limulus Amebocyte Lysate Test (LAL test) is widely used in in vitro quality control assays for endotoxin detection, although it presents some ethical...

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Main Authors: Antonella Di Paolo, Rosario Liberti, Lucia Anzalone, Claudia Colabella, Andrea Felici, Giulio Severi, Monica Cagiola
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Veterinary Sciences
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Online Access:https://www.mdpi.com/2306-7381/11/12/673
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author Antonella Di Paolo
Rosario Liberti
Lucia Anzalone
Claudia Colabella
Andrea Felici
Giulio Severi
Monica Cagiola
author_facet Antonella Di Paolo
Rosario Liberti
Lucia Anzalone
Claudia Colabella
Andrea Felici
Giulio Severi
Monica Cagiola
author_sort Antonella Di Paolo
collection DOAJ
description Background: Veterinary autogenous vaccines, similar to all injectable pharmaceutical products, must be tested to assess endotoxin concentrations. The Limulus Amebocyte Lysate Test (LAL test) is widely used in in vitro quality control assays for endotoxin detection, although it presents some ethical issues related to the production of reagents and is also characterized by a low specificity due to other contaminants that can activate the reaction. For all these reasons, a new recombinant factor C LAL test was developed. Aim: In this study, we described the comparison between two LAL test methods for in vitro quality control of veterinary autogenous vaccines, with the aim of evaluating the most suitable method and establishing an endotoxin concentration range for two different matrices. Methods: Two hundred batches of two different vaccine matrices were tested using the kinetic chromogenic LAL test and recombinant factor C endotoxin detection assay commercial kits. Results and Conclusions: Statistical analysis conducted after the validation of the recombinant factor C test exhibited a statistically significant correlation between the two methods and for both vaccine matrices, suggesting that the animal-free assay can be used as a routine quality control test for veterinary autogenous vaccines.
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series Veterinary Sciences
spelling doaj-art-ae5fee4a10ba4cf0ab5282aa6a8c32c22025-08-20T02:39:42ZengMDPI AGVeterinary Sciences2306-73812024-12-01111267310.3390/vetsci11120673Recombinant Factor C as an In Vitro Assay for the Residual Pathogenicity Evaluation of Veterinary Autogenous VaccinesAntonella Di Paolo0Rosario Liberti1Lucia Anzalone2Claudia Colabella3Andrea Felici4Giulio Severi5Monica Cagiola6Istituto Zooprofilattico Sperimentale Dell’Umbria e Delle Marche “Togo Rosati”, via G. Salvemini 1, 06126 Perugia, ItalyIstituto Zooprofilattico Sperimentale Dell’Umbria e Delle Marche “Togo Rosati”, via G. Salvemini 1, 06126 Perugia, ItalyIstituto Zooprofilattico Sperimentale Dell’Umbria e Delle Marche “Togo Rosati”, via G. Salvemini 1, 06126 Perugia, ItalyIstituto Zooprofilattico Sperimentale Dell’Umbria e Delle Marche “Togo Rosati”, via G. Salvemini 1, 06126 Perugia, ItalyIstituto Zooprofilattico Sperimentale Dell’Umbria e Delle Marche “Togo Rosati”, via G. Salvemini 1, 06126 Perugia, ItalyIstituto Zooprofilattico Sperimentale Dell’Umbria e Delle Marche “Togo Rosati”, via G. Salvemini 1, 06126 Perugia, ItalyIstituto Zooprofilattico Sperimentale Dell’Umbria e Delle Marche “Togo Rosati”, via G. Salvemini 1, 06126 Perugia, ItalyBackground: Veterinary autogenous vaccines, similar to all injectable pharmaceutical products, must be tested to assess endotoxin concentrations. The Limulus Amebocyte Lysate Test (LAL test) is widely used in in vitro quality control assays for endotoxin detection, although it presents some ethical issues related to the production of reagents and is also characterized by a low specificity due to other contaminants that can activate the reaction. For all these reasons, a new recombinant factor C LAL test was developed. Aim: In this study, we described the comparison between two LAL test methods for in vitro quality control of veterinary autogenous vaccines, with the aim of evaluating the most suitable method and establishing an endotoxin concentration range for two different matrices. Methods: Two hundred batches of two different vaccine matrices were tested using the kinetic chromogenic LAL test and recombinant factor C endotoxin detection assay commercial kits. Results and Conclusions: Statistical analysis conducted after the validation of the recombinant factor C test exhibited a statistically significant correlation between the two methods and for both vaccine matrices, suggesting that the animal-free assay can be used as a routine quality control test for veterinary autogenous vaccines.https://www.mdpi.com/2306-7381/11/12/673bacterial endotoxin testingrecombinant factor CLAL testveterinary autogenous vaccine
spellingShingle Antonella Di Paolo
Rosario Liberti
Lucia Anzalone
Claudia Colabella
Andrea Felici
Giulio Severi
Monica Cagiola
Recombinant Factor C as an In Vitro Assay for the Residual Pathogenicity Evaluation of Veterinary Autogenous Vaccines
Veterinary Sciences
bacterial endotoxin testing
recombinant factor C
LAL test
veterinary autogenous vaccine
title Recombinant Factor C as an In Vitro Assay for the Residual Pathogenicity Evaluation of Veterinary Autogenous Vaccines
title_full Recombinant Factor C as an In Vitro Assay for the Residual Pathogenicity Evaluation of Veterinary Autogenous Vaccines
title_fullStr Recombinant Factor C as an In Vitro Assay for the Residual Pathogenicity Evaluation of Veterinary Autogenous Vaccines
title_full_unstemmed Recombinant Factor C as an In Vitro Assay for the Residual Pathogenicity Evaluation of Veterinary Autogenous Vaccines
title_short Recombinant Factor C as an In Vitro Assay for the Residual Pathogenicity Evaluation of Veterinary Autogenous Vaccines
title_sort recombinant factor c as an in vitro assay for the residual pathogenicity evaluation of veterinary autogenous vaccines
topic bacterial endotoxin testing
recombinant factor C
LAL test
veterinary autogenous vaccine
url https://www.mdpi.com/2306-7381/11/12/673
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