Abdulaziz I. Alzarea ¹ Hassan Alhassan ¹ Sami Alzarea ¹ Abdullah S. Alanazi ¹ Fahad O. Alotaibi ² Omar Alsaidam ¹ Mohammed Bakkari ^3*

• ¹ Jouf University, Sakakah, Al Jawf, Saudi Arabia
• ² Forensic Medical Services Center, Riyadh, Saudi Arabia
• ³ Faculty of Pharmacy, Jazan University, Jizan, Saudi Arabia

Paragonimus Westermani is the etiological agent of Paragonimiasis and is mainly transmitted through animals by eating raw or undercooked crustaceans. Currently, no vaccines are available against paragonomiasis. The current research was focused on the identification of vaccine targets using Cysteine Proteases of P. Westermani. The ProtParam bioinformatics tool predicted that the target protein comprising 325 amino acids, with a Molecular weight of 36.11641 kilo Dalton, Theoretical pI, Instability index of 4.96, Aliphatic index30.97 and GRAVY -0.396, physiochemical properties represent that the target proteins are stable, water-soluble. The immunoinformatics analysis predicted that the target proteins is antigenic with a 0.4937 probable antigenic value, the AllerTOP v. 2.0 server predicted that the target is non-allergic. The Cysteine Proteases were used for B-cell epitopes prediction and predicted four linear epitopes using the IEDB tool, the predicted epitopes were used for T-cell epitopes prediction using different MHC alleles. The epitopes were filtered through antigenicity analysis and only four antigenic epitopes were shortlisted for vaccine construction. The epitopes were joined by a GPGPG linker and attached with β-defensin adjuvant to make multi epitopes vaccine construct. The vaccine was analyzed through Immunoinformatics and found that the model vaccine is antigenic, non-allergic, and water-soluble. The docking results show that the model vaccine has a good binding affinity with the immune cell receptors thus it can properly activate the immune system, moreover the Cimmune simulator also unveils that the model vaccine can activate the immune system. The overall immunoinformatics, docking, biophysics, and c immune simulation analysis suggested that the vaccine construct can generate the humoral and cellular immune response and can tackle the pathogenesis of the target parasite.