In Silico Design of a Novel Antibody-Drug Conjugate Targeting Mesothelin in Pancreatic Cancer

Antibody-drug conjugates (ADCs) represent a promising new class of therapeutics thatcombine the specificity of immunotherapy with the potency of chemotherapy, offering a targetedapproach to cancer treatment, including pancreatic cancer[۱], [۲]. This research focuses on the designof an ADC that targets Mesothelin (MSLN), a surface antigen overexpressed in pancreatic tumor cells.In this study, we aimed to design a mesothelin directed antibody conjugate composed of ahumanized single-chain variable fragment (scFv) and GrB using precise in silico techniques.The challenge lies in the phenomenon of ‘shedding’, where proteolytic enzymes cleave various aminoacid sequences, preventing the ADC from binding to the mesothelin on the cancer cell surface[۳]. Toovercome this, we utilized the findings of Liu, X et al., to retrieve the sequence of a humanized singlechainvariable fragment (scFv), known as ۱۵B۶ scFv, which recognizes the residual mesothelin postshedding[۴].We then conjugated ۱۵B۶ to granzyme B (GrB) to design our ADC, employing precise in silicomethodologies[۵]. Four distinct linker peptides were used for the conjugation of ۱۵B۶ to GrB, and the۳D structures of these antibody conjugates were predicted using I-TASSER.The conjugate whose linker peptide least affected the structural conformation of ۱۵B۶ and GrB waschosen. We also compared the solubility and melting temperature of the selected conjugate with thoseof ۱۵B۶ and GrB, and evaluated its physicochemical properties and flexibility.Finally, we compared the binding capacity and the dissociation constant (Kd) of the selected conjugateto MSLN with those of ۱۵B۶, and identified the residues contributing to antigen binding using LigPlot+software.However in vivo and in vitro investigation is required to determine the stability, feasibility andeffectiveness of such construct, in silico techniques, such as those employed in this study, could beutilized for the early development of immune based therapeutics.

کلیدواژه ها:

Antibody-Drug Conjugate ، Mesothelin ، Pancreatic Cancer ، In Silico Design ، Granzyme B