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Updated: Jun 12, 2026
Magnetic Fluorescent Bead-Based Dual-Reporter Flow Analysis of PDL1-Vaxx Peptide Vaccine-Induced Antibody Blockade of the PD-1/PD-L1 Interaction
Published on: July 7, 2023
Engineering and characterization of a novel PD-L1/VEGF bispecific antibody with enhanced VEGF-neutralizing capacity
Qiaoshuang Chen1, Tongyu Hang1, Dan Mao1
1State Key Laboratory of Macromolecular Drugs and Large-Scale Preparation, School of Pharmaceutical Sciences, Cixi Biomedical Research Institute, Wenzhou Medical University, Wenzhou, China; State Key Laboratory of Macromolecular Drugs and Large-Scale Preparation, School of Pharmaceutical Sciences and Food Engineering, Liaocheng University, Liaocheng, China; State Key Laboratory of Macromolecular Drugs and Large-Scale Preparation, NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antibodies, Shanghai Zhangjiang Biotechnology Co., Ltd, Shanghai, China.
Abstract:
Combined blockade of PD-L1 and VEGF represents a promising therapeutic strategy for cancer, yet enhancing anti-VEGF efficacy remains challenging. Here, we engineered a novel bispecific antibody, PLVBP, by fusing a novel high-affinity anti-PD-L1 antibody (T0004) with four VEGF binding domains derived from VEGFR1 D2 (domain II). PLVBP was efficiently expressed in mammalian cells and purified via a three-step chromatography process, achieving over 98% purity. Comprehensive structural characterization confirmed its homogeneity, stability, and preserved secondary structure. Surface plasmon resonance (SPR) analysis revealed that PLVBP binds to PD-L1 and VEGF165 with sub-nanomolar affinities. Functional assays demonstrated that PLVBP maintained potent PD-L1 binding while exhibiting significantly enhanced VEGF binding affinity and capacity compared to bevacizumab. Furthermore, in cell-based assays, PLVBP effectively suppressed VEGF-induced HUVEC proliferation and migration, confirming its VEGF-neutralizing activity. Furthermore, PLVBP simultaneously engaged both PD-L1 and VEGF, validating its dual-targeting capability. Preliminary stability assessments indicated that PLVBP largely maintained its physicochemical and functional integrity during long-term storage in non-optimized buffer. These findings highlight PLVBP as a promising bispecific therapeutic candidate with enhanced avidity-driven VEGF capture, providing a strong foundation for further preclinical development.
