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

Toxins

Benjamin Messham¹  Wai Wan Vivian Tam¹  D. Andrew James¹  Lisa Szymkowicz¹  Jeffery Gerard¹ 

[1] MS/NMR Center, Analytical Sciences, Sanofi Pasteur Ltd., Toronto, ON M2R 3T4, Canada;

关键词: Bordetella pertussis;    adenylate cyclase toxin;    dermonecrotic toxin;    tracheal cytotoxin;    pertussis toxin;    quantitative LC-MS;   

DOI  :  10.3390/toxins13110763

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown