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Updated: Aug 16, 2026
A GPC3-targeting Bispecific Antibody, GPC3-S-Fab, with Potent Cytotoxicity
Published on: July 12, 2018
[Monoclonal antibody against G3BP: preparation, characterization and its application in analysis of human tumors]
Jun-yu Ning1, Jiang-feng You, Fei Pei
1Department of Pathology, Health Science Center, Peking University, Beijing 100083, China.
Objective:
To better understand the molecular mechanism of tumorigenesis and progression, the monoclonal antibody against G3BP (Ras-GAP SH3 binding protein), which serves as an important downstream effector of Ras signaling, was prepared, characterized and utilized in analysis of various human tumors.
Methods:
By using the prokaryotic expression vector pGEX-5X1, GST-G3BP fusion protein was expressed in E. coli BL21 under induction of IPTG. Purified GST-G3BP fusion protein was used to immunize BALB/c mice. The monoclonal antibody against G3BP was produced through conventional hybridoma method and characterized by ELISA, Western blot and immunohistochemical staining.
Results:
A hybridoma cell line secreting anti-G3BP IgG1 subtype antibody was obtained. Western blot and competitive inhibition assay showed that the antibody was G3BP-specific. Immunohistochemical staining demonstrated that G3BP was over-expressed in formalin-fixed and paraffin-embedded tissues of some human tumors, such as lung cancer, colon cancer, gastric cancer and breast cancer. In breast cancer specimens, the degree of G3BP expression correlated positively with the presence of lymph node metastasis and c-erbB2 expression.
Conclusions:
The G3BP-specific monoclonal antibody derived from recombination protein can be used in ELISA, Western blot and immunohistochemical assay. It may provide an important tool in analysis of G3BP in in vitro and in vivo experiments. Besides, G3BP may serve as another prognostic marker for breast cancer.
Insights
A new monoclonal antibody targeting G3BP (Ras-GAP SH3 binding protein) was developed. This antibody is specific for G3BP and shows its overexpression in various cancers, potentially serving as a prognostic marker for breast cancer.
Area of Science:
- Molecular biology
- Oncology
- Immunology
Background:
- Ras signaling pathway is crucial in tumorigenesis.
- G3BP (Ras-GAP SH3 binding protein) is a key downstream effector of Ras.
- Understanding G3BP's role requires specific molecular tools.
Purpose of the Study:
- To develop and characterize a monoclonal antibody against G3BP.
- To analyze G3BP expression in human tumors.
- To evaluate G3BP as a potential biomarker.
Main Methods:
- Prokaryotic expression of GST-G3BP fusion protein.
- Hybridoma technology for monoclonal antibody production.
- ELISA, Western blot, and immunohistochemistry for characterization and analysis.
Main Results:
- A G3BP-specific monoclonal antibody (IgG1 subtype) was successfully generated.
- G3BP was found to be overexpressed in lung, colon, gastric, and breast cancers.
- G3BP expression correlated with lymph node metastasis and c-erbB2 in breast cancer.
Conclusions:
- The developed anti-G3BP antibody is suitable for various assays (ELISA, Western blot, IHC).
- This antibody is a valuable tool for studying G3BP in vitro and in vivo.
- G3BP shows potential as a prognostic marker for breast cancer.
