Host-informed molecular engineering improves antigen quality and enables integrated manufacturing of a trivalent subunit vaccine [Ribo-seq]

We analyzed total RNA and ribosome footprints collected from a K. phaffii strain secreting the VP8-P[6] serotype of the rotavirus antigen and compared against a strain secreting the VP8-P[4] serotype for reference. Overall design: K. phaffii strains expressing either VP8-P[4] or VP8-P[6] were analyzed for total mRNA expression and ribosome protected mRNA quantification. All strains were sampled following biomass accumulation in glycerol-containing medium for 24 hr followed by induction of protein expression by growth in methanol-containing medium for an additional 16 hr.