CENTRO DI RIFERIMENTO ONCOLOGICO
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MONOCLONAL ANTIBODY - 213 anti BCR CRO Aviano researchers have developed a monoclonal antibody, named 213, recognizing an epitope situated across the FWR3 and CDR3 domains of the k light chain of the B Cell Receptor. This antibody has been successfully used in Flow Cytometry toward SH9 cells (spontaneous lymphoblastoid cell line, IgG+ Vk3+, IgM−, EBV+). 213 MAB has also been tested for its ability to mediate Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC). The results indicated that it induced an 8% of lysis in SH9 cells when incubated with peripheral blood mononuclear cells isolated from healthy donors. It is also reactive in ELISA assay and in Western Blot. In particular, in Western Blot, it recognizes the cryoprecipitates isolated from HCV positive patients with Type II mixed Cryoglobulinemia. We have previously demonstrated that the B-cell receptor (BCR) repertoire expressed by type II mixed cryoglobulinemia (II-MC)- associated clonal B-cell proliferations and by HCV-associated non-Hodgkin’s lymphoma (NHL) is not random, with V1–69, V3–7, V4–59 variable heavy (VH)- and still more variable κ (VK)3–20 and VK3–15 light (VL)-chain genes the most represented. We produced some murine MABs, using an Id prototype [a single-chain variable fragment (scFv) antibody], as antigen, which mimicked the VH/VL sequence of a tumor sample obtained from a patient with a B-cell lymphoma, a II-MC disorder, and an HCV infection (De Re, V. et al. 2000, Int. J Cancer 87: 211– Efficient digestion of monoclonal antibody 213 by immobilized Ficin 216). enzyme. Analysis of various experimental fractions by non-reducing SDS-PAGE (12%). Each well was loaded with 2 µl of protein.
Antibody Overview 1. Clone name: 213 2. Specificity: human B Cell Receptor k light chain (VK3–20, VK3–15) 3. Species/Isotype: Murine (IgG2b isotype) 4. This antibody can be used for the following applications and corresponding conditions: _X_ Flow Cytometry _X_ ELISA: detection __x__ _X_ Western Blot: detection __x__ Screening on Western blot. Analysis of Id-scFv (lane 2), a human IgG-K+ (lane 3), human IgG from diffferent isotypes (IgG1, lane 4; IgG2, lane 5; IgG3, lane 6), a polyclonal human IgG (lane 7), and two cryoprecipitates from two II-MC subjects (lane 8 and lane 9) followed by a goat antimouse IgG-HRP. Reactivity toward the Id-scFv used as antigen, and toward human IgG and II-MC-associated cryoprecipitates.
5. What is the best positive tissue/cell line to use in testing this antibody? SH9 cell line or ScFv recombinant protein 6. Publications: De Re V. et al. Annals of the New York Academy of Sciences, 1173: 152–160
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More Information Valli De Re, PhD Department of Molecular Oncology and Translational Medicine (DOMERT) vdere@cro.it +39-0434-659-672/675