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Updated: Jun 1, 2026
A GPC3-targeting Bispecific Antibody, GPC3-S-Fab, with Potent Cytotoxicity
Published on: July 12, 2018
An active murine-human chimeric Fab antibody derived from Escherichia coli, potential therapy against over-expressing
Jianfei Huang1, Jie Liang, Qi Tang
1Department of Pathology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu, China.
Abstract:
Many recombinant murine monoclonal antibodies (mAbs) were studied under pre-clinical or clinical development and became one of the most prolific drug classes in oncology. Vascular endothelial growth factors receptor 2 (VEGFR2) has been implicated to play an important role in tumors. We have established a murine anti-VEGFR2 mAb. To reduce the shortcoming of the mAb, a murine-human chimeric Fab (cFab) named FA8H1 was constructed with gene engineering techniques and expressed as a soluble and functional protein in Escherichia coli Top10F'. Several immunological methods were used to characterize the cFab, including ELISA, affinity and kinetics assay, IP, IF, FACS, and IHC. The results illuminated that cFab maintained the specificity for the VEGFR2 antigen. The active cFab also effectively identified VEGFR2 over-expressing cells in a number of archived human cancer tissues, compared to its parental antibody. The FA8H1 provided the basis for potential therapy research against over-expressing VEGFR2 human solid tumors.
Insights
Researchers engineered a chimeric antibody fragment (cFab) called FA8H1 to target vascular endothelial growth factor receptor 2 (VEGFR2). This new antibody fragment shows promise for developing therapies against human solid tumors overexpressing VEGFR2.
Area of Science:
- Oncology
- Immunology
- Biotechnology
Background:
- Recombinant monoclonal antibodies (mAbs) are a significant class of oncology drugs.
- Vascular endothelial growth factors receptor 2 (VEGFR2) is crucial in tumor development.
- A murine anti-VEGFR2 mAb was developed but had limitations.
Purpose of the Study:
- To engineer a chimeric antibody fragment (cFab) to overcome the limitations of the parental mAb.
- To characterize the functionality and specificity of the novel cFab, FA8H1.
- To assess the potential of FA8H1 for therapeutic research against VEGFR2-overexpressing tumors.
Main Methods:
- Gene engineering techniques were used to construct the murine-human chimeric Fab (cFab) FA8H1.
- FA8H1 was expressed as a soluble protein in Escherichia coli.
- Immunological assays including ELISA, affinity/kinetics assays, immunoprecipitation (IP), immunofluorescence (IF), flow cytometry (FACS), and immunohistochemistry (IHC) were employed for characterization.
Main Results:
- The cFab FA8H1 demonstrated maintained specificity for the VEGFR2 antigen.
- FA8H1 effectively identified VEGFR2-overexpressing cells in archived human cancer tissues.
- The chimeric fragment showed comparable or improved performance over the parental antibody.
Conclusions:
- The engineered FA8H1 cFab is a functional and specific binder of VEGFR2.
- FA8H1 shows potential for identifying and targeting VEGFR2-overexpressing human solid tumors.
- This work lays the foundation for further therapeutic development using FA8H1.
