title: Production and Characterization of New Anti-Human CD20 Monoclonal Antibody.
abstract: The B-cell CD20 antigen is one of the most reliable surface targets in immunotherapy of B lymphoma. In this project, we studied the production and characterization of a new monoclonal antibody against chimeric human CD20 extra loops (hCD20 exl). The results showed that clone C12H, IgG2/k isotype reacted with the antigen in ELISA and immunoblot. The Kd value was found to be 2×10(-9)M and flow cytometry results showed that 99.9% and 99.7% of the Daudi and Raji cells respectively were stained with C12H monoclonal antibody (mab) but not with Jurkat cell lines. It also effectively competed with Rituximab, thus, the staining of the Daudi and Raji cell lines was reduced to 55.9% and 40.5% of cells respectively. Based on the high affinity reaction of C12H mab and appropriate reactivity of C12H mab with the native antigen on the surface of Raji and Daudi cells in flow cytometry, it was concluded that development and evaluation of C12H mab could be a beneficial candidate for further application in genetically engineered monoclonal antibody.
authors.affiliation: Applied Biotechnology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
authors.forename: Mahdi
authors.initials: M
authors.lastname: Fasihi-Ramandi
authors.affiliation: Applied Microbiology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
authors.forename: Jafar
authors.initials: J
authors.lastname: Amani
authors.affiliation: Department of Agricultural Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
authors.forename: AliHatef
authors.initials: A
authors.lastname: Salmanian
authors.affiliation: Immunology Department, Tarbiat Modares University, Tehran, Iran.
authors.forename: SeyedMohammad
authors.initials: S
authors.lastname: Moazzeni
authors.affiliation: Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran.
authors.forename: Kazem
authors.initials: K
authors.lastname: Ahmadi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D018951:Antigens, CD20 / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D006825:Hybridomas; D015151:Immunoblotting; D007074:Immunoglobulin G; D007155:Immunologic Factors / Q000096:biosynthesis*; D008214:Lymphocytes; D016393:Lymphoma, B-Cell / Q000188:drug therapy*; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D000911:Antibodies, Monoclonal; D018951:Antigens, CD20; D007074:Immunoglobulin G; D007155:Immunologic Factors
keywords: CD20; Immunotherapy; Lymphoma; Monoclonal antibody