TY - CONF A1 - Dippong, Martin A1 - Flemig, Sabine A1 - Hoffmann, Katrin A1 - Kunte, Hans-Jörg A1 - Schenk, J. A. A1 - Resch-Genger, Ute A1 - Schneider, Rudolf A1 - Kuhne, Maren T1 - Flow Cytometry (FACS) for characterization and sorting of murine B cells to advance development of monoclonal antibodies T2 - Trends in Diagnostics CY - Tübingen, Germany DA - 2013-10-07 PY - 2013 AN - OPUS4-29241 LA - deu AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER - TY - CONF A1 - Dippong, Martin A1 - Gürnth-Marschner, C. A1 - Kirschbaum, M. A1 - Duschl, C. A1 - Schneider, Rudolf A1 - Kuhne, Maren A1 - Hoffmann, Katrin T1 - Fraunhofer Life Science Day N2 - Their high affinity, selectivity and specificity make monoclonal antibodies (mAbs) very important tools in research, diagnostics and therapy. The production of mAbs is routinely performed by hybridoma technique. Hybridomas are generated by fusing antibody-producing B cells with indefinitely proliferating myeloma cells. Both, cell fusion and the indispensable identification and isolation of the desired antibody-producing hybridoma clone is still far from being trivial. We describe a novel method that should help to optimize the cell fusion, and the screening for antibody-secreting hapten-specific hybridoma cells by using fluorescence activated cell sorting and single-cell fusion. Hybridoma cells specific for a hapten were incubated with a hapten-peroxidase conjugate (hapten-HRP), which was subsequently incubated with a fluorophore-labeled polyclonal anti-peroxidase antibody (anti-HRP-Alexa Fluor® 488). To characterize the expression of membrane-bound immunoglobulin G (IgG) a fluorophore-labeled anti-mouse IgG antibody (anti-IgG-Alexa Fluor® 647) was used. Successful staining was verified by confocal laser scanning microscopy (CLSM). We show that it is possible to specifically label hapten-specific hybridoma cells. It should also be possible to use this labelling approach for the isolation of hapten-specific B cells from the spleen and use these cells for single-cell fusion. Parallelized cell fusion chips were developed to increase fusion efficiencies. T2 - Fraunhofer Life Science Day 2015 CY - Leipzig, Germany DA - 03.07.2015 KW - Single-cell fusion KW - Hybridoma KW - Monoclonal antibodies PY - 2015 AN - OPUS4-37872 LA - eng AD - Bundesanstalt fuer Materialforschung und -pruefung (BAM), Berlin, Germany ER -