In silico design and evaluation of scFv-LukS-PV as a novel immunotoxin for AML therapy

Acute myeloid leukemia (AML) remains a difficult hematologic malignancy that requires more selective therapeutic strategies. Here, we present the in silico design and comprehensive characterization of a recombinant immunotoxin composed of an anti-CD33 single-chain variable fragment (scFv) fused to the LukS-PV pore-forming toxin. Structural modeling and refinement confirmed correct immunoglobulin-domain folding, preserved LukS-PV architecture, and high stereochemical quality. Physicochemical and mRNA analyses indicated favorable stability, solubility, and strong translational compatibility with Escherichia coli. Protein - protein docking predicted specific and high-affinity binding of the scFv to CD33, while molecular-dynamics simulations supported the overall structural stability and functional flexibility of the fusion construct. Codon optimization yielded a high CAI and balanced GC content, suggesting efficient bacterial expression. Immunogenicity analyses predicted non-allergenic and nontoxic properties. Together, these computational findings identify the LukS-PV - anti-CD33 fusion as a promising candidate for targeted AML therapy and establish a framework for subsequent experimental validation.