TY - CHAP A1 - Holzlöhner, Pamela A1 - Schliebs, Erik A1 - Maier, Natalia A1 - Füner, Jonas A1 - Micheel, Burkhard A1 - Heilmann, Katja T1 - Production of monoclonal camelid antibodies by means of hybridoma technology T2 - The journal of immunology Y1 - 2013 SN - 0022-1767 VL - 190 PB - American Assoc. of Immunologists CY - Bethesda ER - TY - GEN A1 - Maier, Natalia A1 - Holzlöhner, Pamela A1 - Hoenow, Anja A1 - Scheunemann, Astrid A1 - Weschke, Daniel A1 - Hanack, Katja T1 - Characterization of monoclonal antibodies generated by in vitro immunization T2 - The journal of immunology N2 - Monoclonal antibodies are highly valuable tools in biomedicine but the generation by hybridoma technology is very time-consuming and elaborate. In order to circumvent the consisting drawbacks an in vitro immunization approach was established by which murine as well as human monoclonal antibodies against a viral coat protein could be developed. The in vitro immunization process was performed by isolation of murine hematopoietic stem cells or human monocytes and an in vitro differentiation into immature dendritic cells. After antigen loading the cells were co-cultivated with naive T and B lymphocytes for three days in order to obtain antigen-specific B lymphocytes in culture, followed by fusion with murine myeloma cells or human/murine heteromyeloma cells. Antigen-specific hybridomas were selected and the generated antibodies were purified and characterized in this study by ELISA, western blot, gene sequencing, affinity measurements. Further the characteristics were compared to a monoclonal antibody against the same target generated by conventional hybridoma technology. Isotype detection revealed a murine IgM and a human IgG4 antibody in comparison to an IgG1 for the conventionally generated antibody. The antibodies derived from in vitro immunization showed indeed a lower affinity for the antigen as compared to the conventionally generated one, which is probably based on the significantly shorter B cell maturation (3 days) during the immunization process. Nevertheless, they were suitable for building up a sandwich based detection system. Therefore, the in vitro immunization approach seems to be a good and particularly fast alternative to conventional hybridoma technology. Y1 - 2016 SN - 0022-1767 SN - 1550-6606 VL - 196 PB - American Assoc. of Immunologists CY - Bethesda ER - TY - JOUR A1 - Kaba, Hani E. J. A1 - Maier, Natalia A1 - Schliebe-Ohler, Nicole A1 - Mayer, Yvonne A1 - Mueller, Peter P. A1 - van den Heuvel, Joop A1 - Schuchhardt, Johannes A1 - Hanack, Katja A1 - Bilitewski, Ursula T1 - Identification of whole pathogenic cells by monoclonal antibodies generated against a specific peptide from an immunogenic cell wall protein JF - Journal of microbiological methods N2 - We selected the immunogenic cell wall beta-(1,3)-glucosyltransferase Bgl2p from Candida albicans as a target protein for the production of antibodies. We identified a unique peptide sequence in the protein and generated monoclonal anti- C. albicans Bgl2p antibodies, which bound in particular to whole C. albicans cells. KW - Candida KW - Diagnostics KW - Flow cytometry KW - Peptides KW - Bgl2p Y1 - 2015 U6 - https://doi.org/10.1016/j.mimet.2014.11.003 SN - 0167-7012 SN - 1872-8359 VL - 108 SP - 61 EP - 69 PB - Elsevier CY - Amsterdam ER - TY - THES A1 - Maier, Natalia T1 - Aufbau eines Testsystems zum Nachweis von Ethylglucuronid (EtG) in Haaren Y1 - 2016 ER - TY - JOUR A1 - Holzlöhner, Pamela A1 - Butze, Monique A1 - Maier, Natalia A1 - Hebel, Nicole A1 - Schliebs, Erik A1 - Micheel, Burkhard A1 - Fuener, Jonas A1 - Heidicke, Gabriele A1 - Hanack, Katja T1 - Generation of murine monoclonal antibodies with specificity against conventional camelid IgG1 and heavy-chain only IgG2/3 JF - Veterinary Immunology and Immunopathology N2 - Camelids possess antibodies with a conventional four-chain structure consisting of two heavy and two light chains (of subclass IgG1) but further they also generate heavy-chain only antibodies (of subclass IgG2 and 3) which are fully functional in antigen binding. In this study subclass-specific murine monoclonal antibodies specific to conventional camelid IgG1 and heavy-chain only IgG2/3 were generated and validated for the use as potent secondary detection reagents. The monoclonal antibodies are able to differentiate between all camelid IgGs, conventional four-chain camelid antibodies (of subclass IgG1) and exclusively heavy chain-only antibodies (of subclasses IgG2 and IgG3). Further these antibodies were used to detect specific immune responses after vaccination of Camelids against bovine corona- and rotavirus strains and different E.coli. and Clostridia - antigens and to identify Erysipelothrix rhusiopathiae infected animals within a herd. The described antibodies are suitable as new secondary agents for the detection of different camelid subclasses and the validation of camelid immune reactions. KW - Camelid antibodies KW - Heavy-chain only antibodies KW - Monoclonal antibodies KW - Secondary antibodies KW - Vaccination KW - Disease monitoring Y1 - 2018 U6 - https://doi.org/10.1016/j.vetimm.2018.01.006 SN - 0165-2427 SN - 1873-2534 VL - 197 SP - 1 EP - 6 PB - Elsevier CY - Amsterdam ER -