Detection of Residual Proteins UL5 and UL29 Using a Targeted Proteomics Approach in HSV529, a Replication-Deficient HSV-2 Vaccine Candidate

Fradin, Manon J.; Pędyczak, Artur; Carpick, Bruce

doi:10.1016/j.xphs.2018.08.018

Cited by 4 publications

(2 citation statements)

References 34 publications

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“…The manufacturer that provided the antigens and the DTaP vaccines will from this point forward be named as 'manufacturer'. All samples were stored at 4 Netherlands), each containing a single amino acid labeled with 13 C and 15 N in lieu of 12 C and 14 N, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…Targeted mass spectrometry is used as a method to quantify proteins of interest with high sensitivity, accuracy and reproducibility [ 11 ]. This method is widely applied in vaccine research, for example, for the quantification of neuramidase and hemagglutinin in influenza vaccine [ 12 ], as well as the detection of residual UL5 and UL29 proteins in a human herpes simplex virus (HSV)-2 viral vaccine candidate [ 13 ]. In addition, quantitative MS-based assays were developed to (i) quantify the amount of PorA and PorB in Neisseria meningitidis outer membrane vesicles (OMVs) adsorbed to aluminum hydroxide [ 14 , 15 ] and (ii) identify and quantify the Bordetella pertussis antigens: Pertussis toxoid (PTd), filamentous hemagglutinin (FHA), pertactin (PRN) and fimbriae 2 (Fim2) in DTaP vaccines [ 16 ].…”

Section: Introductionmentioning

confidence: 99%

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Mass Spectrometry-Based Quantification of the Antigens in Aluminum Hydroxide-Adjuvanted Diphtheria-Tetanus-Acellular-Pertussis Combination Vaccines

et al. 2022

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Vaccines undergo stringent batch-release testing, most often including in-vivo assays for potency. For combination vaccines, such as diphtheria-tetanus-pertussis (DTaP), chemical modification induced by formaldehyde inactivation, as well as adsorption to aluminum-based adjuvants, complicates antigen-specific in-vitro analysis. Here, a mass spectrometric method was developed that allows the identification and quantitation of DTaP antigens in a combination vaccine. Isotopically labeled, antigen-specific internal standard peptides were employed that permitted absolute quantitation of their antigen-derived peptide counterparts and, consequently, the individual antigens. We evaluated the applicability of the method on monovalent non-adjuvanted antigens, on final vaccine lots and on experimental vaccine batches, where certain antigens were omitted from the drug product. Apart from the applicability for final batch release, we demonstrated the suitability of the approach for in-process control monitoring. The peptide quantification method facilitates antigen-specific identification and quantification of combination vaccines in a single assay. This may contribute, as part of the consistency approach, to a reduction in the number of animal tests required for vaccine-batch release.

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Section: Methodsmentioning

confidence: 99%