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

We analyzed the transcriptomes of K. phaffii strains secreting the VP8-P[6] serotype of the rotavirus antigen, and compared against null strains as well as strains secreting the VP8-P[4] or VP8-P[8] serotype in order to diagnose bottlenecks limiting robust secretion of P[6]. Overall design: Multiple replicates and clones of K. phaffii strains expressing either VP8-P[4], VP8-P[6], or VP8-P[8] as well as null (non-expressing) strains were analyzed for RNA expression. All strains were sampled following biomass accumulation in glycerol-containing medium for 24 hr following by induction of protein expression by growth in methanol-containing medium for an additional 24 hr.