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Development and Functional Characterization of Murine Tolerogenic Dendritic Cells
Published on: May 18, 2018
Development and characterization of a novel mouse anti-canine oncostatin M receptor beta monoclonal antibody
Yuxin Zheng1, Zheng Fan2, Jing Zhang3
1College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, China; Laboratory of Protein Engineering and Vaccines, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China.
Abstract:
Oncostatin M receptor beta (OSMRβ) mediates signaling of Oncostatin M (OSM) and interleukine-31 (IL-31), two key cytokines involved in many important biological processes including inflammation and cancer progression. More importantly, OSMRβ might be a potential biomarker and therapeutic target for some diseases, such as inflammatory bowel disease, pruritus and ovarian cancer. In this study, soluble recombinant canine OSMRβ (cOSMRβ) was experimentally expressed as a native antigen to develop an effective cOSMRβ-specific monoclonal antibody (mAb), 2O2, using hybridoma technology. It was demonstrated that 2O2 is able to detect OSMRβ expressed on cell surface using immunofluorescence assay (IFA) and flow cytometry (FACS). This mAb exhibits very high binding affinity to cOSMRβ with the KD and half-maximal effective concentration (EC50) values of 2.49 nM and 96.96 ng/ml, respectively. Meanwhile, it didn't show any cross-relativities with feline OSMRβ (fOSMRβ) and human OSMRβ (hOSMRβ). Moreover, we determined the binding epitope of 2O2, which localizes in the domain VI (DVI, amino acids 623-734) of cOSMRβ. In conclusion, this novel mAb, 2O2, can be used in immunoassays, including IFA, FACS and enzyme-linked immunosorbent assay (ELISA) to facilitate studies in dogs.
Insights
A new monoclonal antibody (mAb), 2O2, specifically targets canine Oncostatin M receptor beta (cOSMRβ). This antibody demonstrates high affinity and can be used in various immunoassays for canine research.
Area of Science:
- Immunology
- Biotechnology
- Veterinary Medicine
Background:
- Oncostatin M receptor beta (OSMRβ) is crucial for signaling by Oncostatin M (OSM) and Interleukin-31 (IL-31).
- OSMRβ plays a role in inflammation and cancer, and is a potential biomarker and therapeutic target for diseases like inflammatory bowel disease, pruritus, and ovarian cancer.
Purpose of the Study:
- To develop a canine OSMRβ (cOSMRβ)-specific monoclonal antibody (mAb) for research applications.
- To characterize the binding affinity, specificity, and epitope of the developed mAb.
Main Methods:
- Soluble recombinant cOSMRβ was expressed as an antigen.
- Hybridoma technology was employed to generate cOSMRβ-specific mAbs.
- Immunofluorescence assay (IFA), flow cytometry (FACS), and binding affinity assays were performed for characterization.
Main Results:
- A novel mAb, designated 2O2, was successfully developed against cOSMRβ.
- mAb 2O2 effectively detected cell surface-expressed cOSMRβ via IFA and FACS.
- The antibody exhibited high binding affinity (KD = 2.49 nM) and specificity, with no cross-reactivity to feline or human OSMRβ.
- The binding epitope was mapped to domain VI (amino acids 623-734) of cOSMRβ.
Conclusions:
- The novel mAb 2O2 is a highly specific and high-affinity reagent for canine OSMRβ.
- mAb 2O2 is suitable for various immunoassays, including IFA, FACS, and ELISA.
- This antibody will facilitate further research on OSMRβ signaling pathways and related diseases in dogs.
