TY - JOUR A1 - Messerschmidt, Katrin A1 - Heilmann, Katja T1 - Toxin-antigen conjugates as selection tools for antibody producing cells JF - Journal of immunological methods N2 - The generation of antibodies with designated specificity requires cost-intensive and time-consuming screening procedures. Here we present a new method by which hybridoma cells can be selected based on the specificity of the produced antibody by the use of antigen-toxin-conjugates thus eliminating the need of a screening procedure. Initial experiments were done with methotrexate as low molecular weight toxin and fluorescein as model antigen. Methotrexate and a methotrexate-fluorescein conjugate were characterized regarding their toxicity. Afterwards the effect of the fluorescein-specific antibody B13-DE1 on the toxicity of the methotrexate-fluorescein conjugate was determined. Finally, first results showed that hybridoma cells that produce fluorescein specific antibodies are able to grow in the presence of fluorescein-toxin-conjugates. KW - Monoclonal antibody KW - Hybridoma technology KW - Selection of antibody producing cells Y1 - 2013 U6 - https://doi.org/10.1016/j.jim.2012.10.010 SN - 0022-1759 VL - 387 IS - 1-2 SP - 167 EP - 172 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Michelchen, Sophia A1 - Micheel, Burkhard A1 - Hanack, Katja T1 - In vitro immunization approach to generate specific murine monoclonal IgG antibodies JF - Journal of immunological methods : JIM N2 - Generating a monoclonal antibody to date is a time intense process that requires immunization of laboratory animals. The transfer of the humoral immune response into in vitro settings enables a shortening of this process and circumvents the necessity of in vivo immunization. However, to orchestrate the complex interplay of dendritic cells, T and B lymphocytes in vitro is very challenging. We therefore aimed for a simplified approach focusing on the protagonist of antibody production: the B lymphocyte. We activated purified murine B lymphocytes alone in vitro by using combinations of antigen and stimuli. We were able to induce a specific antibody response within ten days of culture against a viral coat protein as model antigen. Antibodies were of both IgM and IgG subclass. The stimulated B lymphocytes were transformed into permanently antibody-producing hybridomas by cell fusion technology. We furthermore used this method to induce a specific antibody response against L. pneumophila in vitro. We thus established a useful and effective in vitro protocol to generate monoclonal antibodies. By overcoming the necessity of in vivo immunization this protocol may be the first step towards a universal strategy to generate antibodies from various species. KW - Monoclonal antibody KW - Hybridoma technology KW - In vitro immunization KW - B cell activation Y1 - 2021 U6 - https://doi.org/10.1016/j.jim.2021.113149 SN - 0022-1759 SN - 1872-7905 VL - 499 PB - Elsevier CY - Amsterdam ER - TY - JOUR A1 - Lütkecosmann, Steffi A1 - Warsinke, Axel A1 - Tschöpe, Winfried A1 - Eichler, Rüdiger A1 - Hanack, Katja T1 - A novel monoclonal antibody suitable for the detection of leukotriene B4 JF - Biochemical and biophysical research communications N2 - Leukotriene B4 as an inflammatory mediator is an important biomarker for different respiratory diseases like asthma, chronic obstructive pulmonary disease or cystic lung fibrosis. Therefore the detection of LTB4 is helpful in the diagnosis of these pulmonary diseases. However, until now its determination in exhaled breath condensates suffers from problems of accuracy. Reasons for that could be improper sample collection and preparation methods of condensates and the lack of consistently assay specificity and reproducibility of the used immunoassay detection system. In this study we describe the development and the characterization of a specific monoclonal antibody (S27BC6) against LTB4, its use as molecular recognition element for the development of an enzyme-linked immunoassay to detect LTB4 and discuss possible future diagnostic applications. KW - Leukotriene B4 KW - Monoclonal antibody KW - Immunosensor KW - Chronic obstructive pulmonary disease (COPD) KW - Hapten Y1 - 2017 U6 - https://doi.org/10.1016/j.bbrc.2016.11.157 SN - 0006-291X SN - 1090-2104 VL - 482 IS - 4 SP - 1054 EP - 1059 PB - Elsevier CY - San Diego ER - TY - JOUR A1 - Luetkecosmann, Steffi A1 - Faupel, Thomas A1 - Porstmann, Silvia A1 - Porstmann, Tomas A1 - Micheel, Burkhard A1 - Hanack, Katja T1 - A cross-reactive monoclonal antibody as universal detection antibody in autoantibody diagnostic assays JF - Clinica chimica acta N2 - Diagnostics of Autoimmune Diseases involve screening of patient samples for containing autoantibodies against various antigens. To ensure quality of diagnostic assays a calibrator is needed in each assay system. Different calibrators as recombinant human monoclonal antibodies as well as chimeric antibodies against the autoantigens of interest are described. A less cost-intensive and also more representative possibility covering different targets on the antigens is the utilization of polyclonal sera from other species. Nevertheless, the detection of human autoantibodies as well as the calibration reagent containing antibodies from other species in one assay constitutes a challenge in terms of assay calibration. We therefore developed a cross-reactive monoclonal antibody which binds human as well as rabbit sera with similar affinities in the nanomolar range. We tested our monoclonal antibody S38CD11B12 successfully in the commercial Serazym (R) Anti-Cardiolipin-beta 2-GPI IgG/IgM assay and could thereby prove the eligibility of S38CD11B12 as detection antibody in autoimmune diagnostic assays using rabbit derived sera as reference material. KW - Monoclonal antibody KW - Detection KW - Autoimmune diagnostics KW - Cross reactivity KW - Assay calibration Y1 - 2019 U6 - https://doi.org/10.1016/j.cca.2019.09.003 SN - 0009-8981 SN - 1873-3492 VL - 499 SP - 87 EP - 92 PB - Elsevier CY - Amsterdam ER -