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AssigneesCornell University, Northwestern University
InventorsMichael Christopher Jewett, Jessica Carol Stark, Matthew P. DeLisa, Thapakorn Jaroentomeechai
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Disclosed are methods, systems, components, and compositions for cell-free synthesis of glycosylated proteins. The glycosylated proteins may be utilized in vaccines, including anti-bacterial vaccines. The glycosylated proteins may include a bacterial polysaccharide conjugated to a carrier, which may be utilized to generate an immune response in an immunized host against the polysaccharide conjugated to the carrier. The glycosylated proteins may be synthesized in cell-free glycoprotein synthesis (CFGpS) systems using prokaryote cell lysates that are enriched in components for glycoprotein synthesis such as oligosaccharyltransferases (OSTs) and lipid-linked oligosaccharides (LLOs) including OSTs and LLOs associated with synthesis of bacterial O antigens.
A method for synthesizing an N-glycosylated carrier protein for a bacterial O-antigen via coordinated transcription, translation, and N-glycosylation in vitro, the method comprising:
A method for cell-free production of a glycoprotein, the method comprising:
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