Cargando…

Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays

Botulinum neurotoxin type-A (BoNTA) is one of the seven different serotypes (A to G) produced by Clostridium botulinum. A stability-indicating size-exclusion chromatography (SEC) method was developed and validated, and the specificity was confirmed by forced degradation study, interference of the ex...

Descripción completa

Detalles Bibliográficos
Autores principales: Xavier, Bruna, Perobelli, Rafaela Ferreira, Walter, Maurício Elesbão, da Silva, Francielle Santos, Dalmora, Sérgio Luiz
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6356430/
https://www.ncbi.nlm.nih.gov/pubmed/30642048
http://dx.doi.org/10.3390/toxins11010035
_version_ 1783391538712150016
author Xavier, Bruna
Perobelli, Rafaela Ferreira
Walter, Maurício Elesbão
da Silva, Francielle Santos
Dalmora, Sérgio Luiz
author_facet Xavier, Bruna
Perobelli, Rafaela Ferreira
Walter, Maurício Elesbão
da Silva, Francielle Santos
Dalmora, Sérgio Luiz
author_sort Xavier, Bruna
collection PubMed
description Botulinum neurotoxin type-A (BoNTA) is one of the seven different serotypes (A to G) produced by Clostridium botulinum. A stability-indicating size-exclusion chromatography (SEC) method was developed and validated, and the specificity was confirmed by forced degradation study, interference of the excipients, and peaks purity. The method was applied to assess the content and high-molecular-weight (HMW) forms of BoNTA in biopharmaceutical products, and the results were compared with those of the LD(50) mouse bioassay, the T−47D cell culture assay, and the reversed-phase chromatography (RPC) method, giving mean values of 0.71% higher, 0.36% lower, and 0.87% higher, respectively. Aggregated forms showed significant effects on cytotoxicity, as well as a decrease in the bioactivity (p < 0.05). The employment of the proposed method in conjunction with the optimized analytical technologies for the analysis of the intact and altered forms of the biotechnology-derived medicines, in the correlation studies, enabled the demonstration of the capability of each one of the methods and allowed for great improvements, thereby assuring their safe and effective use.
format Online
Article
Text
id pubmed-6356430
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-63564302019-02-05 Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays Xavier, Bruna Perobelli, Rafaela Ferreira Walter, Maurício Elesbão da Silva, Francielle Santos Dalmora, Sérgio Luiz Toxins (Basel) Article Botulinum neurotoxin type-A (BoNTA) is one of the seven different serotypes (A to G) produced by Clostridium botulinum. A stability-indicating size-exclusion chromatography (SEC) method was developed and validated, and the specificity was confirmed by forced degradation study, interference of the excipients, and peaks purity. The method was applied to assess the content and high-molecular-weight (HMW) forms of BoNTA in biopharmaceutical products, and the results were compared with those of the LD(50) mouse bioassay, the T−47D cell culture assay, and the reversed-phase chromatography (RPC) method, giving mean values of 0.71% higher, 0.36% lower, and 0.87% higher, respectively. Aggregated forms showed significant effects on cytotoxicity, as well as a decrease in the bioactivity (p < 0.05). The employment of the proposed method in conjunction with the optimized analytical technologies for the analysis of the intact and altered forms of the biotechnology-derived medicines, in the correlation studies, enabled the demonstration of the capability of each one of the methods and allowed for great improvements, thereby assuring their safe and effective use. MDPI 2019-01-12 /pmc/articles/PMC6356430/ /pubmed/30642048 http://dx.doi.org/10.3390/toxins11010035 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Xavier, Bruna
Perobelli, Rafaela Ferreira
Walter, Maurício Elesbão
da Silva, Francielle Santos
Dalmora, Sérgio Luiz
Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays
title Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays
title_full Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays
title_fullStr Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays
title_full_unstemmed Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays
title_short Content/Potency Assessment of Botulinum Neurotoxin Type-A by Validated Liquid Chromatography Methods and Bioassays
title_sort content/potency assessment of botulinum neurotoxin type-a by validated liquid chromatography methods and bioassays
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6356430/
https://www.ncbi.nlm.nih.gov/pubmed/30642048
http://dx.doi.org/10.3390/toxins11010035
work_keys_str_mv AT xavierbruna contentpotencyassessmentofbotulinumneurotoxintypeabyvalidatedliquidchromatographymethodsandbioassays
AT perobellirafaelaferreira contentpotencyassessmentofbotulinumneurotoxintypeabyvalidatedliquidchromatographymethodsandbioassays
AT waltermauricioelesbao contentpotencyassessmentofbotulinumneurotoxintypeabyvalidatedliquidchromatographymethodsandbioassays
AT dasilvafranciellesantos contentpotencyassessmentofbotulinumneurotoxintypeabyvalidatedliquidchromatographymethodsandbioassays
AT dalmorasergioluiz contentpotencyassessmentofbotulinumneurotoxintypeabyvalidatedliquidchromatographymethodsandbioassays