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Bioconjugate vaccines' synthesis in prokaryotic cell lysates

A cell lysate, cell-free technology, used in vaccines, intact cells/viruses/DNA/RNA components, bacterial antigen components, etc., and can solve problems such as no proof

Pending Publication Date: 2021-11-12
NORTHWEST UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, PGCT is limited by: i) the length of the in vivo process development timeline; and ii) FDA-approved carrier proteins (such as toxins from Clostridium tetani and Corynebacterium diptheriae) are not yet available. Demonstrating the fact that it is compatible with N-linked glycosylation in living E. coli

Method used

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  • Bioconjugate vaccines' synthesis in prokaryotic cell lysates
  • Bioconjugate vaccines' synthesis in prokaryotic cell lysates
  • Bioconjugate vaccines' synthesis in prokaryotic cell lysates

Examples

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preparation example Construction

[0114] The in vitro bioconjugate vaccine preparation method disclosed in the present invention has demonstrated advantages in rapid, modular and portable vaccine formulation and preparation over existing methods. This system addresses the limitations of existing preparation methods, making it an attractive alternative or complementary strategy for antimicrobial vaccine preparation. Given the growing healthcare problem caused by antibiotic-resistant bacterial infections, this method has the potential to be very valuable for the development, preparation and distribution of new vaccines against a wide variety of pathogenic bacterial strains. Advantages of the methods of the present disclosure include: on-demand expression of bioconjugate vaccines; shaping of novel bioconjugate vaccine candidates; shaping of novel bioconjugate vaccine production pathways; distribution of bioconjugate vaccines to resource-poor settings.

[0115] In conclusion, we disclose the first prokaryotic cell...

Embodiment approach 1

[0119] Embodiment 1. A method of synthesizing an N-glycosylated recombinant protein carrier that can optionally be used as a bioconjugate immunogen or a vaccine, the method comprising performing the synthesis of the recombinant protein carrier Coordinated transcription, translation and N-glycosylation, thereby providing said N-glycosylated recombinant protein carrier, said recombinant protein carrier can be used as a bioconjugate immunogen or vaccine, wherein said N-glycosylated recombinant The protein carrier comprises: (i) a consensus sequence (optionally inserted in said protein carrier) N-X-S / T, where X can be any natural or unnatural amino acid except proline; and (ii) derived from at least one At least one antigenic polysaccharide of bacteria N-linked to said recombinant protein carrier, wherein said at least one antigenic polysaccharide is optionally at least one bacterial O-antigen, optionally from Escherichia coli or Francisella tularensis and the bioconjugate vaccine...

Embodiment approach 2

[0120] Embodiment 2. The method of embodiment 1, wherein the carrier protein is an engineered variant of E. coli maltose binding protein (MBP).

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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...

Claims

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Application Information

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IPC IPC(8): C12P21/02C12P19/18
CPCC12P21/02C12Y204/01C12P21/005A61K39/08A61K39/0208A61K39/0258A61K2039/6037A61K39/00A61K2039/6068A61P31/04A61K2039/53Y02A50/30C07K14/245C07K14/195A61K39/02C07K14/285C07K14/22A61K39/39
Inventor M·C·杰维特J·C·斯塔克M·P·德里萨T·亚罗恩托麦查
Owner NORTHWEST UNIV
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