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Updated: Feb 26, 2026
Generation of Murine Monoclonal Antibodies by Hybridoma Technology
Published on: January 2, 2017
Production of a Mouse Monoclonal Antibody Against Mortalin by Whole Cell Immunization
Marzieh Rezaei1,2, Abbas Ghaderi2
11 Department of Immunology, School of Medicine, Shiraz University of Medical Sciences , Shiraz, Iran .
Abstract:
Pancreatic carcinoma is the fourth leading cause of cancer death and is characterized by early invasion and metastasis. Advances in molecular biology directed new strategies in targeted therapy using monoclonal antibodies. To identify new biomarkers, we generated a panel of monoclonal antibodies against the newly established cell line, Faraz-ICR, from a patient with acinar cell carcinoma. After immunization of BALB/c female mice with Faraz-ICR cell line and fusion of splenocytes with SP2/0 myeloma cell line, high reactive hybridoma producing antibodies to Faraz-ICR were detected using enzyme-linked immunosorbent assay, immunofluorescence staining and flow cytometry. Western blot and two-dimensional immunoblot were used for further characterization of the targets antibodies. Among high reactive clones, the reactivity of 1C11 clone was assessed with other epithelial tumors. The isotype of the antibody was revealed to be IgM, and the antibody reacted to a protein with a molecular weight of about 70 kDa in Western blot analysis. To further characterization of the target antigen, immunoproteome of the Faraz-ICR cell line was performed. By liquid chromatography-mass spectrometry (LC-MS) analysis, we identified that the target of 1C11 clone was HSP70. In conclusion, pancreatic cancer is a fatal malignancy with no reliable biomarker for early screening and diagnosis. In this study, by establishing a pancreatic cell line, a panel of monoclonal antibodies was generated aiming to explore specific or associated cancer targets. We then introduced 1C11 monoclonal antibody that can specifically recognize mortalin as a main tumor marker and may serve as a new tool for diagnostic kit and therapeutic strategies targeting this molecule.
Insights
Researchers developed a new monoclonal antibody, 1C11, targeting HSP70 (Heat Shock Protein 70) in pancreatic cancer. This antibody shows potential as a diagnostic biomarker and therapeutic target for this deadly disease.
Area of Science:
- Oncology
- Immunology
- Molecular Biology
Background:
- Pancreatic carcinoma is a leading cause of cancer mortality, marked by early invasion and metastasis.
- Current diagnostic and therapeutic strategies for pancreatic cancer are limited.
- Monoclonal antibodies offer promising avenues for targeted cancer therapy.
Purpose of the Study:
- To generate novel monoclonal antibodies for identifying new pancreatic cancer biomarkers.
- To characterize the target antigen recognized by the generated antibodies.
- To explore the potential of identified biomarkers in pancreatic cancer diagnostics and therapeutics.
Main Methods:
- Established a new pancreatic acinar cell carcinoma cell line (Faraz-ICR).
- Generated and screened monoclonal antibodies using ELISA, immunofluorescence, and flow cytometry.
- Characterized antibody targets via Western blot, 2D-immunoblot, and LC-MS analysis.
- Identified the target antigen as HSP70 (Heat Shock Protein 70).
Main Results:
- A panel of monoclonal antibodies was generated against the Faraz-ICR cell line.
- The 1C11 antibody clone demonstrated specific reactivity.
- LC-MS analysis identified the target antigen of the 1C11 antibody as HSP70.
- The 1C11 antibody recognizes mortalin, a potential tumor marker.
Conclusions:
- HSP70 (Heat Shock Protein 70), recognized by the 1C11 antibody, is identified as a potential tumor marker for pancreatic cancer.
- The 1C11 monoclonal antibody may serve as a valuable tool for developing diagnostic kits.
- This antibody presents potential for novel therapeutic strategies targeting pancreatic cancer.
