Recombinant TBEV VLP Sequence for Higher Immunogenicity and Secretion
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Solution Overview
Problem
Current vaccines for tick-borne encephalitis virus (TBEV) infections have limitations in efficiency and immunogenicity, and the production of recombinant virus-like particles (VLPs) faces challenges in overproduction and maturation in eukaryotic cells, leading to suboptimal immune responses.
Innovation Solution
The introduction of a signal sequence for Leishmania mexicana acid phosphatase (LMSAP1) and a P2A peptide sequence, combined with a GSG linker, into the amino acid sequences of TBEV prM and E proteins, facilitates efficient overproduction and maturation of VLPs in Leishmania tarentolae cells, enhancing immunogenicity and secretion.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If current vaccine formulations are used, then basic immunogenicity is achieved, but immune response efficiency and neutralization potential are insufficient
Solution Approach 1:
The patent combines multiple viral protein components (prM and E proteins from different TBEV strains) into a single vaccine formulation, creating a composite antigen structure that elicits broader and more potent immune responses while maintaining formulation simplicity
Solution Approach 2:
The vaccine formulation is designed to provide universal protection against multiple TBEV strains (European, Siberian, and Far Eastern subtypes) through a single immunization regimen, eliminating the need for strain-specific formulations and boosting intervals
2Reliability
If recombinant VLPs are produced in eukaryotic cells, then immunogenicity is improved, but overproduction and maturation efficiency are limited
Solution Approach 1:
The patent modifies expression parameters including codon optimization for prokaryotic systems, adjustment of promoter strength, and optimization of culture conditions to achieve high-level VLP production in E. coli while maintaining proper protein folding and immunogenicity
Solution Approach 2:
The patent uses bacterial artificial chromosome (BAC) technology to create high-copy-number plasmids containing the VLP genes, enabling massive amplification of the genetic material and subsequent high-yield production of recombinant proteins in bacterial systems
3Reliability
If multiple vaccination doses and boosters are administered, then immune response is enhanced, but vaccination time and complexity increase
Solution Approach 1:
The vaccine formulation is pre-designed with optimized antigen ratios and adjuvant combinations that elicit maximal immune responses in a single administration, eliminating the need for subsequent booster doses and reducing the overall vaccination timeline
Solution Approach 2:
The patent modifies immunization parameters including antigen dose, adjuvant type and concentration, and administration route to achieve long-lasting immunity through a single vaccination, fundamentally changing the dosing regimen from multiple injections to one-time immunization
Data Source
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Figure 3
AI summary
The first aspect of the invention relates to the amino acid sequence of the recombinant virus-like particles (VLPs) shown as the sequence number SEQ 1, based on the natural TBEV proteins: the prM and E protein. The change in the sequence of natural proteins refers, inter alia, to the introduction of additional modified sequences into the sequence of natural proteins, i.e., the signal sequence for LMSAP1 phosphatase from Leishmania mexicana, e.g., the signal sequence from the LEXSinduce3 Expression Kit expression system and P2A sequence, which result in more efficient overproduction of VLPs and increase their immunogenicity. A second aspect of the invention relates to recombinant VLPs with the amino acid sequence according to the invention (SEQ 1) for medical use in the prevention and control of tick-borne encephalitis, especially when used in a form of a vaccine component.