# Multi-epitope vaccine design against Clostridioides difficile using the ABC-type transport system protein

## TL;DR

This paper presents a new multi-epitope vaccine design targeting Clostridioides difficile using an ABC-type transport system protein to prevent infections.

## Contribution

A novel multi-epitope vaccine construct with TLR3 adjuvant is proposed and validated for immunogenicity and structural stability.

## Key findings

- The vaccine construct showed excellent structural stability and strong receptor binding affinity.
- Immune simulations indicated elevated IFN-γ and IL-2 levels, suggesting strong B- and T-cell responses.
- The construct was confirmed to be antigenic, non-allergenic, and physicochemically stable.

## Abstract

Clostridioides difficile is a major cause of antibiotic-associated nosocomial diarrhoea with increasing global incidence and nearly
one million annual cases reported in India. Therefore, it is of interest to design a multi-epitope vaccine design against Clostridioides
difficile the ABC-type transport system protein. Hence, B-cell, CTL and HTL epitopes were predicted using BepiPred-2.0, NetCTL 1.2 and
NetMHCIIpan 4.0. Thus, a 66-amino-acid epitope construct with AAY and GPGPG spacers for the TLR3 adjuvant to enhance immunogenicity was
reported for further consideration. The construct was validated for antigenicity, non-allergenicity and physicochemical stability.
Structural modelling, molecular docking with TLR3 (PDB ID: 2A0Z) and 100-ns molecular dynamics simulations showed excellent structural
stability (RMSD ~0.1 Å) and receptor binding affinity, while immune simulations via C-ImmSim indicated strong B- and T-cell responses
with elevated IFN-γ and IL-2 levels, supporting its potential as a safe, effective, and targeted prophylactic against C. difficile
infections.