DEVELOPMENT AND CHARACTERIZATION OF AN ATP-BASED METHOD FOR DETERMINING THE NUMBER OF VIABLE UNITS IN RECOMBINANT BACILLE CALMETTE, GUERIN VACCINE

Abstract

The only licensed vaccine against human pulmonary tuberculosis is BCG; it is a live, attenuated vaccine whose viability is conventionally determined by the CFU assay. Due to the testing time requirements and the variability in the colony counts, the CFU assay has drawbacks for BCG/rBCG vaccine manufacturing in-process and quality control product release testing. By utilizing ATPs usefulness as a marker for cell viability, a faster assay was developed and characterized for validation feasibility. The ATP assay is an easy-to-perform alternative and was shown to correlate to the CFU. Overall, the data indicate that while the ATP is a rapid alternative method for quantifying the viability of BCG/rBCG, validation of the assay will have to be sample-specific.