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Updated: Apr 14, 2026
Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood
Published on: February 6, 2018
Generation and Characterization of a New Monoclonal Antibody Against CXCL4
Jing Gao1, Mingyuan Wu, Jin Gao
11 Laboratory of Regeneromics, School of Pharmacy, School of Agriculture and Biology, Shanghai Jiao Tong University , Shanghai, China .
Abstract:
CXCL4 plays important roles in numerous disease processes, which makes the CXCL4 signaling pathway a potential therapeutic target. In this study, we aimed to develop a neutralizing antibody against both human and mouse CXCL4. Rats were immunized with recombinant human CXCL4 (rhCXCL4). Hybridoma clones were created by fusion of the immunized rat spleen cells with mouse myeloma SP2/0 cells and screened using recombinant mouse CXCL4 (rmCXCL4) and rhCXCL4. The CXCL4 monoclonal antibody (CXCL4 MAb) produced by the 16D6-3 hybridoma clone was sequenced and characterized by Western blot and Biacore assays. It recognized both human and mouse CXCL4 with high affinity and neutralized the effect of rhCXCL4 in vitro. Thus, the antibody may be used in the studies of CXCL4 in murine disease models and as a template in the antibody humanization for clinical developments.
Insights
Researchers developed a neutralizing antibody targeting CXCL4, a key molecule in disease. This antibody effectively neutralizes both human and mouse CXCL4, offering potential for therapeutic development and research in disease models.
Area of Science:
- Immunology
- Molecular Biology
- Drug Discovery
Background:
- Chemokine (C-X-C motif) ligand 4 (CXCL4) is implicated in various disease processes.
- The CXCL4 signaling pathway presents a potential therapeutic target for numerous conditions.
Purpose of the Study:
- To develop a novel monoclonal antibody (MAb) capable of neutralizing both human and mouse CXCL4.
- To characterize the affinity and neutralizing capacity of the developed CXCL4 MAb for potential therapeutic applications.
Main Methods:
- Immunization of rats with recombinant human CXCL4 (rhCXCL4).
- Generation and screening of hybridoma clones using recombinant mouse CXCL4 (rmCXCL4) and rhCXCL4.
- Sequencing and characterization of the lead antibody clone (16D6-3) using Western blot and Biacore assays.
Main Results:
- A high-affinity monoclonal antibody (CXCL4 MAb) was successfully developed from the 16D6-3 hybridoma clone.
- The antibody demonstrated cross-reactivity, recognizing both human and mouse CXCL4.
- In vitro studies confirmed the neutralizing activity of the CXCL4 MAb against rhCXCL4.
Conclusions:
- The developed CXCL4 MAb is a valuable tool for studying CXCL4 in murine disease models.
- This antibody serves as a promising template for antibody humanization strategies aimed at clinical development.
