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Development and Characterization of a New Antipeptide Monoclonal Antibody Directed to Human CD20 Antigen
Mahdi Habibi-Anbouhi1, Kayhan Azadmanesh2, Mahdi Behdani3
11 National Cell Bank of Iran, Pasteur Institute of Iran . Tehran, Iran .
Abstract:
The rapid expansion of immunotherapeutic approaches for treatment of various diseases, including cancers, has been greatly facilitated by the invention of new generation of antibodies. Clinical studies have indicated that anti-CD20 mAb-based therapies represent an effective treatment for various diseases with overexpression of CD20 on their cell surface, such as non-Hodgkin's lymphoma, hemolytic anemia, as well as autoimmune diseases like rheumatoid arthritis. Technically, due to a short extra membrane domain, the recombinant CD20 protein is a difficult antigen to raise immune responses. In search for new monoclonal antibodies, the authors used an antigenic polypeptide, which yielded numbers of new binders that may lead to production of anti-CD20 antibodies, with improved diagnostic or clinical attributes. Mice were immunized with extra membrane loop of human CD20 (exCD20) polypeptide. The exCD20 antigen showed a desired immune response and was able to develop a monoclonal antibody, 3B4C10, which reacted well with peptide antigen as well as native antigen on the surface of Raji B-cell line. The antibody 3B4C10 with a balanced K(on) and K(off) may be applicable in the construction of affinity columns or beads for isolation and purification of CD20-positive cells and cancer stem cells.
Insights
Researchers developed a new monoclonal antibody, 3B4C10, targeting the CD20 protein. This antibody, derived from an extracellular polypeptide, shows potential for improved diagnostics and therapies for CD20-expressing cancers and autoimmune diseases.
Area of Science:
- Immunology
- Biotechnology
- Oncology
Background:
- Immunotherapies, particularly anti-CD20 monoclonal antibodies (mAbs), are effective for cancers like non-Hodgkin's lymphoma and autoimmune diseases.
- The CD20 protein's short extracellular domain presents challenges for generating effective immune responses and developing new antibodies.
- There is a need for novel anti-CD20 antibodies with enhanced diagnostic and clinical utility.
Purpose of the Study:
- To develop novel monoclonal antibodies targeting the CD20 antigen.
- To overcome challenges associated with using the full CD20 protein as an antigen.
- To identify antibodies with potential for improved diagnostic or therapeutic applications.
Main Methods:
- Immunization of mice with an antigenic polypeptide representing the extracellular loop of human CD20 (exCD20).
- Screening for monoclonal antibodies that bind to both the peptide antigen and native CD20 on B-cells.
- Characterization of antibody binding kinetics (K(on) and K(off)).
Main Results:
- Successful generation of a new monoclonal antibody, designated 3B4C10.
- 3B4C10 demonstrated reactivity with both the exCD20 peptide and native CD20 on the Raji B-cell line.
- The antibody exhibited balanced binding kinetics (K(on)/K(off)), suggesting high affinity and stability.
Conclusions:
- The exCD20 polypeptide is an effective antigen for generating anti-CD20 antibodies.
- The novel antibody 3B4C10 shows promise for applications in cell isolation and purification.
- Potential uses include affinity columns or beads for isolating CD20-positive cells, including cancer stem cells.
