Surface display of antigens on gram-negative outer membrane vesicles
Inventors
SALVERDA, Merijn Louis Marten • Van Der Ley, Peter André
Assignees
Nederlanden Volksgezondheid Welzijn en Sport VWS • Intravacc BV
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Publication Number
US-10639280-B2
Publication Date
2020-05-05
Expiration Date
Abstract
The present invention relates to vaccine compositions based on Gram-negative outer membrane vesicles displaying antigens of pathogens expressed as part of a fusion protein comprising N-terminal parts of surface expressed lipoproteins of Gram-negative bacteria, and use of such compositions in vaccination. The invention further relates to the fusion lipoproteins comprising N-terminal parts of surface expressed lipoproteins of Gram-negative bacteria and antigens of pathogens fused thereto, DNA constructs and bacterial host cells for expressing these fusion lipoproteins and to methods for producing outer membrane vesicles displaying the fusion lipoproteins.
Core Innovation
The invention relates to a fusion lipoprotein comprising an N-terminal fusion partner and a C-terminal fusion partner. The N-terminal fusion partner comprises, in N- to C-terminal order, a lipidated N-terminal cysteine and a tether of a surface exposed lipoprotein of a Gram-negative bacterium, and optionally further comprises a stretch of at least 5 contiguous amino acids located C-terminally of the tether. The N-terminal fusion partner causes presentation of the fusion lipoprotein on the extracellular outer membrane surface of a Gram-negative bacterium upon expression therein.
The C-terminal fusion partner comprises at least one epitope of an antigen associated with an infectious disease and/or a tumour. The overall amino acid sequence of the fusion lipoprotein does not occur in nature, and the C-terminal fusion partner does not comprise an amino acid sequence of at least 10 contiguous amino acids from the surface exposed lipoprotein from which the sequence of the N-terminal fusion partner originates.
The described concept enables outer-membrane surface display of antigen epitopes using engineered fusion lipoproteins. It includes examples where pathogen antigens are redirected to the outer membrane surface and used to elicit antibody responses.
Claims Coverage
The document provides one independent claim defining a fusion lipoprotein with specified N-terminal and C-terminal fusion partners, including constraints on contiguous sequence derivation from a Gram-negative surface exposed lipoprotein and a requirement that the overall amino acid sequence does not occur in nature.
N-terminal lipidated cysteine tether for extracellular outer membrane presentation
An N-terminal fusion partner comprising a lipidated N-terminal cysteine and a tether of a surface exposed lipoprotein of a Gram-negative bacterium, wherein the N-terminal fusion partner causes presentation of the fusion lipoprotein on the extracellular outer membrane surface of a Gram-negative bacterium upon expression therein.
Optional contiguous residues C-terminally of the tether
Optionally, the N-terminal fusion partner further comprises a stretch of at least 5 contiguous amino acids located C-terminally of the tether of a surface exposed lipoprotein of a Gram-negative bacterium.
C-terminal antigen epitope for infectious disease and/or tumour
A C-terminal fusion partner comprising at least one epitope of an antigen associated with an infectious disease and/or a tumour.
C-terminal avoids contiguous sequence from the N-terminal source lipoprotein
The C-terminal fusion partner does not comprise an amino acid sequence of at least 10 contiguous amino acids from the surface exposed lipoprotein from which the sequence of the N-terminal fusion partner originates.
Non-natural overall fusion lipoprotein sequence
The amino acid sequence of the fusion lipoprotein does not occur in nature.
Independent claim covers a non-natural fusion lipoprotein that is presented on the extracellular outer membrane surface of a Gram-negative bacterium through an N-terminal lipidated cysteine plus surface-exposed lipoprotein tether, while the C-terminal partner displays antigen epitopes associated with infectious disease and/or a tumour, without including a contiguous sequence region derived from the N-terminal source lipoprotein.
Stated Advantages
enables presentation of the fusion lipoprotein on the extracellular outer membrane surface of a Gram-negative bacterium upon expression therein
provides at least one epitope of an antigen associated with an infectious disease and/or a tumour on the fusion lipoprotein
Documented Applications
prophylaxis and treatment using outer membrane vesicle (OMV) formulations bearing surface-exposed fusion lipoproteins that include antigen epitopes
immunogenicity indicated by strong mouse antibody responses to surface-exposed antigen fusions
surface display and immunogenicity of non-OspA antigens (e.g., OspC fragments and RmpM) when fused to the OMV surface display system
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