Development and validation of capillary electrophoresis sodium dodecyl sulfate (CE-SDS) method for purity analysis of pertussis toxin, filamentous haemagglutinin and pertactin antigens

We report here the development and validation of CE-SDS method for purity analysis of Acellular Pertussis vaccine components viz. purified Pertussis toxin (PTx), purified Filamentous haemagglutinin (FHA), and Pertactin antigen (PRN). The method was found to be specific and showed excellent linearity at a concentration range of 15.62 mu g/mL-1000 mu g/mL for purified PTx, 31.25 mu g/mL-1000 mu g/mL for purified FHA, and 3.9 mu g/ mL-1000 mu g/mL for PRN antigen. Method reported limit of quantification (LOQ) 31.25 mu g/mL, 62.5 mu g/mL, and 7.8 mu g/mL for purified PTx, FHA, and PRN respectively. Method precision (repeatability and intermediate precision) for purity and molecular weight determination in product matrix was below 10% for all three proteins. Method comparability studies were performed with SDS-PAGE. CE-SDS demonstrated corroborating results with SDS-PAGE for the estimation of purity and molecular weight analysis. However, CE-SDS method exhibited better resolution capabilities for resolving all the sub-unit peaks of PTx and isoforms of purified FHA. CE-SDS method also demonstrated stability indicating potential and thus fits its intended purpose as an effective analytical tool for quality control of acellular pertussis-based vaccines.

Automated summary

We have developed and validated a CE-SDS method for analyzing the purity of components in acellular Pertussis vaccine. The method demonstrated specificity and excellent linearity over a concentration range. CE-SDS showed better resolution capabilities compared to SDS-PAGE.