Construction, expression, and characterization of a multi-epitope protein based on envelope glycoproteins B, C, and D of Varicellovirus bovinealpha1/5 in Escherichia coli.

Engineering an Innovative Chimeric Multi-Epitope RNA-Based Vaccine Against Neonatal Calf Diarrhea Pathogens (Bovine Coronavirus, Bovine Rotavirus, and Escherichia coli K99): An In Silico-Based Analysis

Neonatal calf diarrhea (NCD) is one of the most important problems of calf breeding across the world. It causes deaths in calves in the first 10 days of their life, and it is mainly caused by Escherichia coli(E. coli), Bovine Rotavirus (BRV) and Bovine Coronavirus (BCoV). The lack of vaccines with consistently high protective efficacy against the main causes of NCD makes disease control highly challenging. The current study aims to design a multi-epitope mRNA-based vaccine targeting the major pathogens responsible for NCD using immunoinformatic tools and molecular modeling approaches. BRV capsid protein VP6, BCoV Spike glycoprotein and E. coli F5 fimbrial protein were used as antigenic proteins to predict potential epitopes. Fifteen selected epitopes were linked with suitable linkers and conjugated with a built-in adjuvant, resulting in the design of a stable, antigenic and non-allergenic vaccine candidate against NCD pathogens. Furthermore, molecular docking analysis shows strong binding affinity between the vaccine candidate and the bovine toll-like receptors TLR2 and TLR4 at low energy and high stability. Based on these findings, the proposed multi-epitope vaccine represents a promising approach for the prevention and control of neonatal calf diarrhea and provides a solid scientific foundation for future experimental studies to validate its efficacy and safety in vivo.

Identi fi cation and characterization of new B cell epitopes on the nucleocapsid protein of bovine coronavirus using monoclonal antibodies

Development and immunological evaluation of a multi-antigen recombinant adenovirus vaccine candidate against PRRSV and PCV2

In this study, a recombinant adenovirus vaccine candidate constructed and evaluated to addressed the widespread clinical problem of co-infections of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Circovirus Type 2 (PCV2). Epidemiological analysis revealed a PRRSV-positive rate of 40.8%, with prevalent strains belonging to Lineage 1 and Lineage 8. Analysis of key amino acid sites in the GP5 protein indicated the presence of highly virulent strain characteristics and widespread wild-type strain variation. Based on these findings and previous studies, we selected multiple PRRSV proteins (GP2–GP5, M, N) from multiple strains and the validated PCV2 Cap protein fused with dominant T/B cell epitopes, and successfully constructed four recombinant adenoviruses using a 2A peptide-linked strategy. These constructs stably expressed the target antigens in HEK293 cells and demonstrated good passage stability. Animal experiments showed that piglets immunized with the recombinant adenoviruses developed high levels of PRRSV- and PCV2-specific antibodies and neutralizing antibodies, and exhibited lymphocyte proliferation and secretion of cytokines including IFN-γ, IL-2, and IL-4, indicating the vaccine simultaneously elicited both humoral and cellular immune responses. Following PRRSV challenge, immunized groups, particularly the multi-antigen combined immunization group, demonstrated significant clinical protection. This study provides a promising vaccine candidate and a theoretical foundation for the coordinated control of PRRSV and PCV2.

Immunoinformatics-Guided Design and In Silico Evaluation of a Multi-Epitope Vaccine Against Influenza A H10N5 and H3N2 Strains Based on Hemagglutinin and Neuraminidase Proteins

Conserved epitope-driven in silico design of a multi-epitope vaccine against the tick-borne wetland virus.

These computational results suggest that the construct has the potential to induce effective immunity against WELV, however further experimental validations in future have to be performed in order to confirm its efficacy against the pathogen.

An Innovative Approach towards Construction, Evaluation, Simulations and Cloning of a Multi-epitope Vaccine against Ebola Virus, through Immunoinformatics-based Study Design

Introduction Ebola virus has been preying on human populations for a long time. But recent outbreaks in Africa with symptoms such as severe haemorrhagic fever urge the immediate need for research on an Ebola vaccine as an initial step. Hence, in this study, an in silico vaccine is designed against the Ebola virus, with epitopes representing all three strains: Zaire, Sudan, and Bundibugyo. Methodologies After initial sequence retrieval, they are prioritized based on physicochemical analyses and other suitable parameters. Epitopes are then predicted from selected sequences and subjected to stringent analyses – antigenicity, allergenicity, toxicity, conservation, solubility, among other parameters. An in silico vaccine is constructed using epitopes, incorporating Beta-defensin 3 at the C-terminus and a His6 Tag at the N-terminus. After a stringent set of analyses, the protein structure is subjected to Secondary and Tertiary structure prediction. After refinement of the structure, validation procedures are performed using PROCHECK, PRO-SA, and Molprobity. Simulation methods such as GROMACS and Immune Simulations, as implemented in C-IMMISIM, are used. Thereafter, docking with TLR4 (in PATCHDOCK software) , interaction analyses of the docking mechanism , by PISA software, Codon Optimization by JCAT and cloning into the pET28a(+) with SnapGene , are achieved. Results After applying stringent analytical methods, epitopes are selected for in silico vaccine construction against Ebola virus strains – Zaire, Sudan, and Bundibugyo. The constructed vaccine is subjected to similar analyses again, yielding a Vaxijen score of 0.6853. Secondary structure prediction by PSIPRED ensures reliable conformity, while tertiary structure prediction by Colab AlphaFold2 further reveals the vaccine structure. After refinement, the validation process reveals that the protein vaccine structure contains over 98% Ramachandran-favoured regions and an ERRAT score of 71%. Docking of the vaccine against the TLR4 immune receptor by PATCHDOCK is achieved, and PISA interaction analyses reveal a binding energy of -14.5kcal/mol. Again, Molecular Dynamics simulations reveal its structural integrity and conformity amid solvation analyses. Then, for C-IMMISIM results, both humoral and cell-mediated immunity are activated, along with memory cell propagation. Again, in JCAT, the CAI score is 0.9572, and a GC content of 52.95% is achieved, indicating high expression potential. Conclusion The immunoinformatics tools applied suggest its conformational integrity, compactness, and safety for further wet-lab research. World -class population coverage of 87.44% , along with approx 70% coverge for West, East and Central Africa , reflects the need for further labotory-based experiments with this vaccine candidate.