Bioconjugate vaccines' synthesis in prokaryotic cell lysates

Abstract

Disclosed are methods, systems, components, and compositions for cell-free synthesis of glycosylated proteins, which 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. Suitable carriers may include but are not limited to Haemophilus influenzae protein D (PD), Neisseria meningitidis porin protein (PorA), Corynebacterium diphtheriae toxin (CRM 197), Clostridium tetani toxin (TT), and Escherichia coli maltose binding protein, and variants thereof.

Description

[0001] Statement Regarding Federally Sponsored Research or Development

[0002] This invention was made with government support under MCB1413563 awarded by the US National Science Foundation. The government has certain rights in this invention.

[0003] Cross references to related patent applications

[0004] This application claims the benefit of priority under 35 U.S.C. §119(e) to U.S. Provisional Patent Application No. 62 / 791,425, filed January 11, 2019, which is incorporated herein by reference in its entirety. Background technique

[0005] The field of the invention relates to the in vitro synthesis of N-glycosylated proteins in prokaryotic cell lysates. In particular, the field of the invention relates to the use of N-glycosylated proteins synthesized in vitro in prokaryotic cell lysates as vaccine conjugates against pathogens such as bacteria.

[0006] Conjugate vaccines are one of the safest and most effective methods for preventing life-threatening bacterial infect...

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