Development of a targeted nanoLC-MS/MS method for quantitation of residual toxins from Bordetella pertussis

Szymkowicz, Lisa; Wilson, Derek J.

doi:10.1016/j.jpba.2020.113395

Cited by 6 publications

(9 citation statements)

References 49 publications

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“…al. [27] describing a PRM workflow for protein toxins, ACT and DNT, on a Q-Exactive HF nanoLC-MS/MS system. For the development of a routine test, several additional requirements are typically considered, including system sensitivity, system reliability, assay throughput, ease of analysis, and LC-MS system compliance requirements, which are governed by good manufacturing practices.…”

Section: Discussionmentioning

confidence: 99%

“…In this work, we developed an in vitro method for detection and quantitation of residual B. pertussis toxins, ACT, DNT, and TCT, using a targeted LC-MS (MRM) method. The residual toxin method described by our group previously [27] was transferred to a tandem quadrupole system to facilitate routine analysis requiring higher throughput and the addition of an external calibration curve for TCT. A fully quantitative method is described for ACT and TCT, while DNT detection is currently suitable as a limit test.…”

Section: Discussionmentioning

confidence: 99%

“…A MRM method was designed for targeted quantitation of ACT, DNT, and TCT. Surrogate peptides for ACT and DNT were leveraged from the previously published Parallel Reaction Monitoring (PRM) method [27], with one sequence for quantitation and a second for specificity (Table 1). Given similar collision-induced fragmentation mechanisms on Orbitrap and Triple Quadrupole mass spectrometers, it was expected that the highest responding fragment ions from PRM experiments on the Q Exactive HF Orbitrap would be applicable to MRM experiments on the Xevo TM TQ-S. A Sequence selected for protein quantitation.…”

Section: Multiple Reaction Monitoring (Mrm) Analysismentioning

confidence: 99%

“…Previously, we reported the development of a targeted nano-LC tandem mass spectrometry (MS/MS) method for quantitation of ACT and DNT residual toxins from B. pertussis [27]. Herein we describe the design and qualification of a higher throughput LC-MS/MS method for detection and quantitation of ACT, DNT, and TCT.…”

Section: Introductionmentioning

confidence: 99%

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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

Szymkowicz¹,

Gerard²,

Messham³

et al. 2021

Toxins

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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.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Multiple Reaction Monitoring (Mrm) Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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

Szymkowicz¹,

Gerard²,

Messham³

et al. 2021

Toxins

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“…Finally, batch release of pertussis vaccines still requires a test for residual dermonecrotic toxin in mice [17]. Recently, the use of liquid chromatography mass spectrometry to quantify dermonecrotic toxin has been proposed as an alternative in vitro method [78].…”

Section: Critical Quality Attributes Of Vaccinesmentioning

confidence: 99%

Overcoming scientific barriers in the transition fromin vivoto non-animal batch testing of human and veterinary vaccines

Biggelaar

Hoefnagel

Vandebriel

et al. 2021

Expert Review of Vaccines

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Introduction: Before release, vaccine batches are assessed for quality to evaluate whether they meet the product specifications. Vaccine batch tests, in particular of inactivated and toxoid vaccines, still largely rely on in vivo methods. Improved vaccine production processes, ethical concerns, and suboptimal performance of some in vivo tests have led to the development of in vitro alternatives.Areas covered: This review describes the scientific constraints that need to be overcome for replacement of in vivo batch tests, as well as potential solutions. Topics include the critical quality attributes of vaccines that require testing, the use of cell-based assays to mimic aspects of in vivo vaccine-induced immune responses, how difficulties with testing adjuvanted vaccines in vitro can be overcome, the use of altered batches to validate new in vitro test methods, and how cooperation between different stakeholders is key to moving the transition forward. Expert opinion: For safety testing, many in vitro alternatives are already available or at an advanced level of development. For potency testing, in vitro alternatives largely comprise immunochemical methods that assess several, but not all critical vaccine properties. One-to-one replacement by in vitro alternatives is not always possible and a combination of methods may be required.

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A highly sensitive method for determination of tacrolimus in peripheral blood mononuclear cells by nano liquid chromatography–high resolution accurate mass spectrometry

Qin

Chen

Wang

et al. 2023

Journal of Chromatography A

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Development of a targeted nanoLC-MS/MS method for quantitation of residual toxins from Bordetella pertussis

Cited by 6 publications

References 49 publications

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

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

Overcoming scientific barriers in the transition fromin vivoto non-animal batch testing of human and veterinary vaccines

A highly sensitive method for determination of tacrolimus in peripheral blood mononuclear cells by nano liquid chromatography–high resolution accurate mass spectrometry

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