Purification design and practice for pertactin, the third component of acellular pertussis vaccine, from Bordetella pertussis

Li, Zenglan; Zhang, Yan; Qi, Wang; Li, Zhengjun; Liu, Yongdong; Zhang, Songping; Ma, Guanghui; Luo, Jian; Su, Zhiguo

doi:10.1016/j.vaccine.2016.06.029

Cited by 10 publications

(3 citation statements)

References 27 publications

(27 reference statements)

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“…These constructs can be expressed in E. coli and spiked into evaluation samples, prior to digestion, to assess method performance metrics such as accuracy and precision across a relevant concentration range. Alternatively, the constructs can be expressed in N 15 enriched media to add a metabolic heavy label into every amide bond in the QconCAT sequences. N 15 heavy labelling would allow the QconCAT to be used as an internal quantitation standard [45] in lieu of AQUA peptide standards for ACT and DNT.…”

Section: Discussionmentioning

confidence: 99%

“…Alternatively, the constructs can be expressed in N 15 enriched media to add a metabolic heavy label into every amide bond in the QconCAT sequences. N 15 heavy labelling would allow the QconCAT to be used as an internal quantitation standard [45] in lieu of AQUA peptide standards for ACT and DNT.…”

Section: Discussionmentioning

confidence: 99%

“…The aP vaccines are typically comprised of protein antigens purified from B. pertussis cultures, including pertussis toxin (PT) and other virulence adhesin proteins such as fimbria, pertactin, and filamentous haemagglutinin [14,15]. Because the antigens are produced from fermentations of the causative agent, there are other toxins that can co-purify; it is important that these additional toxins are controlled during development and refinement of purification processes for aP vaccines.…”

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: Introductionmentioning

confidence: 99%

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