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

ABSTRACT: 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].

ORGANISM(S): Komagataella phaffii

PROVIDER: GSE159325 | GEO | 2021/05/05

REPOSITORIES: GEO

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

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

Project description:This SuperSeries is composed of the SubSeries listed below.

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Scalable, methanol-free manufacturing of the SARS-CoV-2 receptor binding domain in engineered Komagataella phaffii

Project description:Prevention of COVID-19 on a global scale will require the continued development of high-volume, low-cost platforms for the manufacturing of vaccines to supply on-going demand. Vaccine candidates based on recombinant protein subunits remain important because they can be manufactured at low costs in existing large-scale production facilities that use microbial hosts like Komagataella phaffii (Pichia pastoris). Here, we report an improved and scalable manufacturing approach for the SARS-CoV-2 spike protein receptor binding domain (RBD); this protein is a key antigen for several reported vaccine candidates. We genetically engineered a manufacturing strain of K. phaffii to obviate the requirement for methanol-induction of the recombinant gene. Methanol-free production improved the secreted titer of the RBD protein by >5x by alleviating protein folding stress. Removal of methanol from the production process enabled scale up to a 1,200 L pre-existing production facility. This engineered strain is now used to produce an RBD-based vaccine antigen that is currently in clinical trials and could be used to produce other variants of RBD as needed for future vaccines.

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

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Streptococcus agalactiae LC-MSMS

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