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Fig 1.

Schematic representation of the study.

Flow chart of the strategy used to design a Multiepitope peptide-based vaccine against the HEV using In-Silico approach.

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Table 1.

Selected epitopes to be used to design multi-epitope vaccine construct.

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Fig 2.

Population coverage analysis.

Schematic representation of global population coverage of the selected epitopes.

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Fig 3.

MHC Class-wise population coverage of the prioritized epitopes.

A) World-class MHC I population coverage. B) World-class MHC II population coverage. C) Class-combined global population coverage graph.

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Fig 4.

Characteristics of designed vaccine construct.

A) Designed vaccine construct consisting of epitopes, adjuvants, and linkers. B) The predicted secondary structure of vaccine sequence. C) The Ramachandran chart depicting 86% residues in its most favoured region.

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Fig 5.

Structural characteristics of the vaccine model.

A) Three-dimensional model for multi-epitope vaccine model. B) physiochemical properties of the vaccine model.

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Table 2.

Binding energy details for the galaxy refined models of the multi-epitope peptides-based vaccine.

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Fig 6.

Mutant model for vaccine generated by introducing cysteine residues (shown in violet).

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Table 3.

Pairs of residues mutated as cysteine.

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Fig 7.

In-silico cloning using E. coli expression system.

Reverse translated vaccine’s DNA sequence as shown in red, expressed in pET-28a (+) vector. GC content for the designed vaccine was 56.23% while the CAI value was 1.0.

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Fig 8.

Multiepitope peptide HEV vaccine docked with the human immune receptor.

TLR3-complex and protein-protein interaction between the chains.

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Table 4.

Energies calculated for TLR-3-complex.

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Fig 9.

Computationally immune simulated host immune response towards Multiepitope peptide HEV vaccine of A) Production of immunoglobulins in response to vaccine model. B) Induced interleukins in response to vaccine model The purpose of Computational immune simulation was to get an immunogenic profile for the designed vaccine [27]. The designed vaccine was observed as inducing primary and secondary immune responses. The production of IgM+IgG was observed at a higher level leading to the production of IgG1 and IgG2 depicting the induction of primary immune response. Production of IgM+IgG was raised up to 700000/ml in the first 15 days, then there is a decline as shown in Fig 9A. IgM and IgG are types of antibodies produced by the immune system in response to an infection or vaccination. It was observed that IFN-g induced was in greater amount while the release of IL-also happened up to 10 days and then start decreasing. IL stands for interleukin, which is a group of cytokines involved in regulating immune responses and inflammation. IFN-g was produced at the higher rate. Its production reached 400000/ml in 5 days and the decrease in its production can be seen after 20 days (Fig 9B). IFN-g refers to interferon-gamma, a type of cytokine that plays a role in immune responses against viral and bacterial infections.

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Fig 10.

Trajectories obtained for simulated complexes (vaccine and receptor).

A) RMSD graph. B) RMSF graph. C) ROG graph. D) B-factor graph.

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