Plant-Produced Influenza VLPs via Modified Hemagglutinin Proteolytic Loop
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Solution Overview
Problem
Current vaccines, such as subunit vaccines, face challenges with poor immunogenicity due to incorrect protein folding and antigen presentation, and often elicit primarily humoral responses, failing to provide effective immunity against influenza viruses, particularly for diseases where whole inactivated virus provides protection.
Innovation Solution
Production of virus-like particles (VLPs) in plants using a nucleic acid with an expression enhancer linked to a modified influenza hemagglutinin (HA) protein, featuring a modified proteolytic loop to reduce or abolish cleavage sites, allowing for enhanced expression and purification of VLPs that can induce a robust immune response.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If subunit vaccines are used, then safety is improved by avoiding whole virus, but immunogenicity deteriorates due to incorrect folding and poor antigen presentation
Solution Approach 1:
The patent creates virus-like particles that copy the native viral structure and organization without containing viral genetic material. The VLPs replicate the authentic viral architecture including proper protein folding and antigen presentation, thereby achieving both safety (no viral genome) and immunogenicity (native-like structure).
Solution Approach 2:
The patent modifies the proteolytic loop of hemagglutinin to reduce or abolish cleavage sites, changing the proteolytic resistance parameter. This modification prevents premature cleavage and maintains proper protein folding and stability, improving both manufacturing precision and vaccine effectiveness while retaining safety.
2Reliability
If subunit vaccines are used, then safety is improved by avoiding whole virus, but immune response effectiveness deteriorates by eliciting primarily humoral responses
Solution Approach 1:
The VLPs copy the native viral structure including surface glycoproteins in their authentic configuration, enabling them to present multiple antigenic determinants simultaneously. This elicits both humoral and cellular immune responses, improving overall immune response effectiveness while maintaining safety through absence of viral genome.
3Productivity
If expression enhancer is added to increase HA protein yield, then productivity is improved, but protein stability may deteriorate due to overexpression
Solution Approach 1:
The patent modifies the proteolytic loop to reduce or abolish cleavage sites, changing the proteolytic resistance parameter. This enhancement of protein stability allows the system to tolerate higher expression levels from the expression enhancer without compromising protein integrity, thereby resolving the contradiction between productivity and stability.
4Stability of the object's composition
If modified proteolytic loop is introduced to reduce cleavage sites, then protein stability is improved, but manufacturing complexity increases
Solution Approach 1:
The patent extracts and modifies only the specific proteolytic loop region of the hemagglutinin gene, leaving the rest of the viral genome structure intact. This targeted modification approach minimizes the complexity increase in the nucleic acid construct while achieving the desired protein stability enhancement.
Data Source
AI summary
A method of producing a virus like particle (VLP) in a plant comprising modified hemagglutinin is provided. The method comprises introducing a nucleic acid comprising a regulatory region active in the plant and operatively linked to a nucleotide sequence encoding a modified influenza hemagglutinin (HA) protein into the plant, or portion of the plant, the modified HA protein comprises a modified proteolytic loop. Followed by incubating the plant or portion of the plant under conditions that permit the expression of the nucleic acids, thereby producing the VLP. The modified proteolytic loop may comprise one or more protease cleavage sites exhibiting reduced or abolished cleavage by a protease. The nucleotide sequence encoding the HA may be selected from the group consisting of B HA, C, H1, H2, H3, H4, H5, H6, H7, H8, H9, H10, H11, H12, H13, H14, H15, and H16. Also described is a virus like particle (VLP) produced by the method, and plants expressing the VLP. The virus like particle (VLP) may comprise plant-specific N-glycans, or modified N-glycans.