This study successfully constructed and characterized a bispecific antibody (anti-EGFR/VEGF-A BsAb) based on the CrossMab/KIH platform. It demonstrated simultaneous targeting of EGFR and VEGF-A, effectively inhibiting EGF/EGFR and VEGF-A/VEGFR2 signaling pathways in ovarian cancer cells, providing experimental evidence for dual-antibody applications in ovarian cancer therapy.
Literature Overview
The article "Physicochemical and biological characterization of a bispecific antibody in a CrossMab/KIH format that targets EGFR and VEGF-A", published in Frontiers in Immunology, reviews the construction of CrossMab/KIH-based bispecific antibodies and their functional validation in ovarian cancer-related signaling pathways. Through genetic engineering, researchers developed a bispecific antibody targeting both EGFR and VEGF-A, systematically evaluating its structural stability, binding capacity, and biological activity under various experimental conditions to establish theoretical foundations for subsequent bioanalytical method development and quality control strategies.
Background Knowledge
Ovarian cancer (Ovarian Cancer, OC) is the deadliest gynecological malignancy, with a 5-year survival rate of only approximately 50.8%. Current treatment primarily involves surgery combined with adjuvant chemotherapy, but 50-70% of patients experience recurrence and develop resistance to existing targeted therapies. EGFR (epidermal growth factor receptor) and VEGF-A (vascular endothelial growth factor A) are highly expressed in OC, promoting cancer cell proliferation and angiogenesis respectively, making them promising therapeutic targets. However, single-target therapies show limited clinical efficacy. Researchers proposed a bispecific antibody strategy targeting both EGFR and VEGF-A to achieve synergistic antitumor effects. The CrossMab/KIH technology combines domain crossover (CrossMab) and heavy chain heterodimer stabilization (KIH) to enhance correct antibody assembly and prevent mispairing. This study investigates the physicochemical properties and biological activity of this dual-target antibody, providing data support for quality control and bioanalytical development.
Research Methods and Experiments
The research team constructed the anti-EGFR/VEGF-A BsAb using public sequences combined with CrossMab/KIH technology. The BsAb was expressed in HEK293 cells and purified through MabSelect™ PrismA and Prism G affinity chromatography. Structural integrity and purity were evaluated via SDS-PAGE, CE-SDS, and SEC-HPLC. ELISA and Biacore assays assessed binding activity and affinity to EGFR and VEGF-A. Functional validation included evaluating inhibition of EGFR and VEGFR2 phosphorylation, as well as VEGF-A secretion, in OC cell lines (CaOV3, SKOV3, OVCAR3, PA-1) and HUVEC cells. Additionally, thermal stability was analyzed under varying temperature conditions.
Key Conclusions and Perspectives
Research Significance and Prospects
This study validates the feasibility of the CrossMab/KIH platform for bispecific antibody construction and demonstrates the anti-EGFR/VEGF-A BsAb's dual-targeting activity in OC and HUVEC models. These findings provide foundational data for developing BsAb-based therapeutics against EGFR and VEGF-A, while offering references for quality attribute evaluation and bioanalytical method development. Future studies should focus on in vivo efficacy testing in animal models to assess antitumor and anti-angiogenic potential, and explore its application in combination targeted therapy strategies.
Conclusion
This study successfully developed a CrossMab/KIH platform-derived bispecific antibody that effectively suppresses EGFR and VEGF-A-mediated signaling in ovarian cancer and endothelial cell models. Research data demonstrate the BsAb's favorable structural stability, binding activity, and thermal resilience, with dual blockade of EGFR and VEGF-A/VEGFR2 pathways suggesting potential synergistic antitumor effects. These results establish a solid foundation for preclinical development and bioanalytical method optimization, offering novel insights into dual-targeting therapies for ovarian cancer.
