Abstract
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.
Highlights
Pertussis disease, commonly known as whooping cough, is a highly infectious respiratory disease caused by the bacterium Bordetella pertussis
Tracheal cytotoxin was detected via nanoLC-MS/MS analysis of B. pertussis supernatant harvest digest
The MS/MS fragmentation pattern observed by high-energy collision induced dissociation (HCD) (Figure 2c) was similar to that obtained by FAB-MS of tracheal cytotoxin (TCT) from B. pertussis [29]
Summary
Commonly known as whooping cough, is a highly infectious respiratory disease caused by the bacterium Bordetella pertussis. The first effective vaccines for prevention of this disease were developed nearly a century ago in the 1930s using inactivated whole-cell B. pertussis bacteria [1,2]. Despite the long history of both the wP and aP vaccines, the disease remains a major public health concern, with nearly 60 thousand deaths (2015) and the largest disease burden among children less than five years in developing countries [6,7]. Acellular vaccines are as efficacious as wP vaccines; the protection provided by acellular vaccines declines faster [10]. This has increased interest in developing new Pertussis vaccines [11,12,13]
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