Cargando…
Design of a Quantitative LC-MS Method for Residual Toxins Adenylate Cyclase Toxin (ACT), Dermonecrotic Toxin (DNT) and Tracheal Cytotoxin (TCT) in Bordetella pertussis Vaccines
The antigens for acellular pertussis vaccines are made up of protein components that are purified directly from Bordetella pertussis (B. pertussis) bacterial fermentation. As such, there are additional B. pertussis toxins that must be monitored as residuals during process optimization. This paper de...
Autores principales: | , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8624556/ https://www.ncbi.nlm.nih.gov/pubmed/34822547 http://dx.doi.org/10.3390/toxins13110763 |
_version_ | 1784606203469365248 |
---|---|
author | Szymkowicz, Lisa Gerard, Jeffery Messham, Benjamin Tam, Wai Wan Vivian James, D. Andrew |
author_facet | Szymkowicz, Lisa Gerard, Jeffery Messham, Benjamin Tam, Wai Wan Vivian James, D. Andrew |
author_sort | Szymkowicz, Lisa |
collection | PubMed |
description | The antigens for acellular pertussis vaccines are made up of protein components that are purified directly from Bordetella pertussis (B. pertussis) bacterial fermentation. As such, there are additional B. pertussis toxins that must be monitored as residuals during process optimization. This paper describes a liquid chromatography mass spectrometry (LC-MS) method for simultaneous analysis of residual protein toxins adenylate cyclase toxin (ACT) and dermonecrotic toxin (DNT), as well as a small molecule glycopeptide, tracheal cytotoxin (TCT) in a Pertussis toxin vaccine antigen. A targeted LC-MS technique called multiple reaction monitoring (MRM) is used for quantitation of ACT and TCT, which have established limits in drug product formulations. However, DNT is currently monitored in an animal test, which does not have an established quantitative threshold. New approaches for DNT testing are discussed, including a novel standard based on concatenated quantitation sequences for ACT and DNT. Collectively, the method represents a “3-in-1” analytical simplification for monitoring process-related residuals during development of B. pertussis vaccines. |
format | Online Article Text |
id | pubmed-8624556 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-86245562021-11-27 Design of a Quantitative LC-MS Method for Residual Toxins Adenylate Cyclase Toxin (ACT), Dermonecrotic Toxin (DNT) and Tracheal Cytotoxin (TCT) in Bordetella pertussis Vaccines Szymkowicz, Lisa Gerard, Jeffery Messham, Benjamin Tam, Wai Wan Vivian James, D. Andrew Toxins (Basel) Article The antigens for acellular pertussis vaccines are made up of protein components that are purified directly from Bordetella pertussis (B. pertussis) bacterial fermentation. As such, there are additional B. pertussis toxins that must be monitored as residuals during process optimization. This paper describes a liquid chromatography mass spectrometry (LC-MS) method for simultaneous analysis of residual protein toxins adenylate cyclase toxin (ACT) and dermonecrotic toxin (DNT), as well as a small molecule glycopeptide, tracheal cytotoxin (TCT) in a Pertussis toxin vaccine antigen. A targeted LC-MS technique called multiple reaction monitoring (MRM) is used for quantitation of ACT and TCT, which have established limits in drug product formulations. However, DNT is currently monitored in an animal test, which does not have an established quantitative threshold. New approaches for DNT testing are discussed, including a novel standard based on concatenated quantitation sequences for ACT and DNT. Collectively, the method represents a “3-in-1” analytical simplification for monitoring process-related residuals during development of B. pertussis vaccines. MDPI 2021-10-28 /pmc/articles/PMC8624556/ /pubmed/34822547 http://dx.doi.org/10.3390/toxins13110763 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Szymkowicz, Lisa Gerard, Jeffery Messham, Benjamin Tam, Wai Wan Vivian James, D. Andrew Design of a Quantitative LC-MS Method for Residual Toxins Adenylate Cyclase Toxin (ACT), Dermonecrotic Toxin (DNT) and Tracheal Cytotoxin (TCT) in Bordetella pertussis Vaccines |
title | Design of a Quantitative LC-MS Method for Residual Toxins Adenylate Cyclase Toxin (ACT), Dermonecrotic Toxin (DNT) and Tracheal Cytotoxin (TCT) in Bordetella pertussis Vaccines |
title_full | Design of a Quantitative LC-MS Method for Residual Toxins Adenylate Cyclase Toxin (ACT), Dermonecrotic Toxin (DNT) and Tracheal Cytotoxin (TCT) in Bordetella pertussis Vaccines |
title_fullStr | Design of a Quantitative LC-MS Method for Residual Toxins Adenylate Cyclase Toxin (ACT), Dermonecrotic Toxin (DNT) and Tracheal Cytotoxin (TCT) in Bordetella pertussis Vaccines |
title_full_unstemmed | Design of a Quantitative LC-MS Method for Residual Toxins Adenylate Cyclase Toxin (ACT), Dermonecrotic Toxin (DNT) and Tracheal Cytotoxin (TCT) in Bordetella pertussis Vaccines |
title_short | Design of a Quantitative LC-MS Method for Residual Toxins Adenylate Cyclase Toxin (ACT), Dermonecrotic Toxin (DNT) and Tracheal Cytotoxin (TCT) in Bordetella pertussis Vaccines |
title_sort | design of a quantitative lc-ms method for residual toxins adenylate cyclase toxin (act), dermonecrotic toxin (dnt) and tracheal cytotoxin (tct) in bordetella pertussis vaccines |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8624556/ https://www.ncbi.nlm.nih.gov/pubmed/34822547 http://dx.doi.org/10.3390/toxins13110763 |
work_keys_str_mv | AT szymkowiczlisa designofaquantitativelcmsmethodforresidualtoxinsadenylatecyclasetoxinactdermonecrotictoxindntandtrachealcytotoxintctinbordetellapertussisvaccines AT gerardjeffery designofaquantitativelcmsmethodforresidualtoxinsadenylatecyclasetoxinactdermonecrotictoxindntandtrachealcytotoxintctinbordetellapertussisvaccines AT messhambenjamin designofaquantitativelcmsmethodforresidualtoxinsadenylatecyclasetoxinactdermonecrotictoxindntandtrachealcytotoxintctinbordetellapertussisvaccines AT tamwaiwanvivian designofaquantitativelcmsmethodforresidualtoxinsadenylatecyclasetoxinactdermonecrotictoxindntandtrachealcytotoxintctinbordetellapertussisvaccines AT jamesdandrew designofaquantitativelcmsmethodforresidualtoxinsadenylatecyclasetoxinactdermonecrotictoxindntandtrachealcytotoxintctinbordetellapertussisvaccines |