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Computational design and validation of a multi-epitope subunit vaccine against the neglected parasite blastocystis sp.

Created on 13 Jul 2026

Authors

Amirtesh Raghuram, Gayathri Mahalingam

Published in

In silico pharmacology. Volume 14. Issue 2. Pages 187. Epub Jul 11, 2026.

Abstract

Blastocystis sp. is a neglected, globally prevalent intestinal parasite infecting over one billion people with no available vaccine. This study describes an immunoinformatics framework for developing a multi-epitope subunit vaccine against α-L-fucosidase from Blastocystis sp. Immunogenic B-cell and T-cell epitopes were identified through systematic prediction and rigorous allergenicity, toxicity, and human homology screening. Epitopes were assembled into a vaccine with human β-defensin-3 as an N-terminal adjuvant. The vaccine displayed favorable physicochemical properties including optimal molecular weight (~ 38 kDa), stability index (< 40) and negative GRAVY score. Population coverage analysis achieved 97.13% global coverage with 24.75 epitope-HLA hits per individual and a PC90 of 14.42. Structural validation of predicted 3D structure using Ramachandran analysis (99.7% residues in favored regions) and ProSA-web evaluation (Z-score: - 5.07) confirmed high-quality tertiary structure. Molecular docking revealed extensive binding with TLR-4, MHC Class I, and MHC Class II receptors, with interface areas of 1379-2198 Å2 and 13-19 hydrogen bonds per complex. VD-MM/GBSA analysis reported favorable binding energies (- 295 to - 370 kcal/mol), while NMA confirmed appropriate dynamic flexibility for biological activity. Immune simulations predicted robust humoral and Th1-biased cellular responses with sustained IgG/IgM antibody production and maintained immune homeostasis. In silico cloning into pET-28a( +) showed optimal codon adaptation (CAI: 0.97) and GC content (53.31%), providing a pathway toward experimental validation of the first Blastocystis vaccine.

PMID:
42438779
Bibliographic data and abstract were imported from PubMed on 13 Jul 2026.

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