CloneID: 3B2-3C9
Antigen Long Description: The original antibody was derived from Epstein-Barr Virus transformed human lymphoblast cells.
Buffer Composition: PBS with 0.02% Proclin 300.
Available Custom Conjugation Options: AP, HRP, Fluorescein, APC, PE, Biotin Type A, Biotin Type B, Streptavidin, FluoroProbes 647H, Atto488, APC/Cy7, PE/Cy7
Uniprot Accession No.: Q9UI17
Specificity Statement: This antibody is specific for the human disialoganglioside GD2, a surface antigen highly expressed on human neuroblastoma cells. The antibody showed high specificity for GD2 without cross-reactivity to other similar gangliosides like GD1a, GM2, and GM3. Surface plasmon resonance and immunostaining techniques confirmed its strong and specific binding to GD2.
Application Notes (Clone): Affinity testing of the original antibody (human IgM) to GD2 via SPR revealed a Kd of 4.5 nM. The antibody effectively labeled GD2-expressing cells in both flow cytometry (FC) and immunofluorescence (IF) assays. ELISA testing across different GD2-expressing cell lines (M14, LLC-MK2, and EL4) showed a high binding affinity for M14 and LLC-MK2 (0.13 ± 0.02 nM and 0.15 ± 0.04 nM, respectively), compared to EL4 (0.93 ± 0.16 nM). A biotinylated version of the antibody was used to stain various melanoma cell lines in an immunohistochemistry (IHC) assay. GD2-positive cells (M14) demonstrated strong staining, while GD2-negative lines (MG1055, RPMI7951, and JS0592) exhibited minimal or no staining, confirming the antibody's specificity. Additionally, the antibody was tested in an IHC panel of 35 normal human tissues to assess cross-reactivity, with most normal tissues showing no significant high-affinity staining, indicating minimal cross-reactivity and specificity toward GD2-expressing cells. The antibody successfully mediated complement-dependent cytotoxicity (CDC) on multiple GD2-positive melanoma cell lines (M14, LF0023, M101, M18, and M10VAC). Furthermore, the IgM version of the antibody exhibited superior CDC activity compared to a human IgG1 isotype-switched version (US8507657B2).
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