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A murine monoclonal antibody raised against human non-small cell carcinoma of the lung
1Center for Molecular Medicine and Immunology, University of Medicine and Denistry of New Jersey, Newark 07103.
Abstract:
Murine monoclonal antibodies (MAbs) reactive with human non-small cell carcinoma of the lung (NSCCL) were produced following immunization with a membrane preparation of Calu-3, a human adenocarcinoma of the lung cell line grown in nude mice. Positive hybrids unreactive with normal liver membranes and human peripheral white blood cells were selected for further testing. One MAb (RS5-4H6) recognized an antigen expressed in a variety of lung and other carcinoma cell lines as detected by flow cytometry. Immunohistology showed a selectivity for normal and neoplastic lung epithelium, as well as other cancers. The antigen was detected by immunohistology in 87% of tumor specimens derived from the lung, breast, colon, kidney, and ovary. Most other normal human tissues were not stained but occasional cells of the stomach, salivary and other glands, as well as kidney tubules were reactive. This MAb is an IgG1. Western blot analysis indicated that the antibody reacts selectively with an antigen greater than 300 kD. The antigen is resistant to neuraminidase treatment and periodate oxidation, but sensitive to pronase treatment, suggesting that the epitope is peptide in nature. This antibody may be potentially useful as a targeting agent for radioimmunodetection and immunoconjugate therapy.
Insights
Researchers developed a novel monoclonal antibody (MAb) targeting a large antigen found in various cancers, including lung carcinoma. This antibody shows potential for cancer detection and therapy.
Area of Science:
- Oncology
- Immunology
- Biochemistry
Background:
- Developing targeted therapies for non-small cell lung carcinoma (NSCCL) requires identifying specific tumor antigens.
- Murine monoclonal antibodies (MAbs) offer specificity for antigen targeting in cancer diagnostics and therapeutics.
Purpose of the Study:
- To produce and characterize MAbs reactive with human NSCCL.
- To evaluate the potential of a specific MAb (RS5-4H6) as a diagnostic and therapeutic agent.
Main Methods:
- Immunization of mice with Calu-3 adenocarcinoma cell line membranes.
- Hybridoma selection based on reactivity with tumor cells and lack of reactivity with normal tissues.
- Flow cytometry and immunohistochemistry for antigen detection.
- Western blot analysis to determine antigen characteristics.
Main Results:
- One MAb, RS5-4H6 (IgG1), recognized an antigen present in various carcinoma cell lines and tumor specimens (87% of lung, breast, colon, kidney, ovary).
- The antigen, >300 kD, showed selectivity for normal and neoplastic lung epithelium and some other tissues.
- Antigen epitope is peptide-based, resistant to neuraminidase and periodate, but sensitive to pronase.
Conclusions:
- MAb RS5-4H6 targets a broadly expressed tumor-associated antigen.
- This MAb demonstrates potential utility for radioimmunodetection and immunoconjugate therapy in various cancers.