Research Information System
EU 7TH INTERNATIONAL CONFERENCE ON HEALTH, ENGINEERING AND APPLIED SCIENCES, Rome, Italy, 1 - 03 March 2025, pp.171, (Summary Text)
Breast cancer is the most common type of cancer worldwide and the second most common in
our country. In recent years, the design of various therapeutic fusion proteins using recombinant
DNA technology for the development of antibody-based drugs in targeted cancer treatment has
become an important field of study. Human Epidermal Growth Factor Receptor-2 (HER2) is
overexpressed in 20-25% of breast cancers. In this study, a single-chain variable fragment of
anti-HER2, a monoclonal antibody targating HER2, which plays a role in the proliferation and
growth of breast cancer cells, was fused with Defensin-β, an antimicrobial peptide known for
its anticancer activity. A single-chain variable fragment fusion protein (Anti-HER2-scFv-
Defensin-β) was designed for use in breast cancer treatment. The 948-base pair long fusion
gene was cloned into the pQE2 expression vector, and expression experiments were performed
in different E. coli strains. It was determined that the highest expression level was achieved in
the E. coli Origami strain. The heterologously expressed Anti-HER2-scFv-Defensin-β protein
was purified and confirmed via SDS-PAGE analysis. Optimization of ultrafiltration and dialysis
methods is ongoing to obtain protein production with high concentration and purity.
Subseqyently, the Anti-HER2-scFv-Defensin-β fusion protein will be applied to cultered breast
cancer cell at different concentrations, and its effectiveness in breast cancer treatment will be
evaluated.
Keywords: Therapeutic proteins and peptides, Cancer, Recombinant DNA Technology