TY - JOUR T1 - In vitro immunization approach to generate specific murine monoclonal IgG antibodies JF - bioRxiv DO - 10.1101/2020.11.24.393728 SP - 2020.11.24.393728 AU - Sophia Michelchen AU - Burkhard Micheel AU - Katja Hanack Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/11/24/2020.11.24.393728.abstract N2 - Generating monoclonal antibodies to date is a time intense process requiring immunization of laboratory animals. The transfer of the humoral immune response into in vitro settings shortens this process and circumvents the necessity of animal immunization. However, orchestrating the complex interplay of immune cells in vitro is very challenging. We aimed for a simplified approach focusing on the protagonist of antibody production: the B lymphocyte. We activated purified murine B lymphocytes in vitro with combinations of antigen and stimuli. Within ten days of culture we induced specific IgM and IgG antibody responses against a viral coat protein. Permanently antibody-producing hybridomas were generated. Furthermore we used this method to induce a specific antibody response against Legionella pneumophila. We thus established an effective protocol to generate monoclonal antibodies in vitro. By overcoming the necessity of in vivo immunization it may be the first step towards a universal strategy to generate antibodies from various species.Competing Interest StatementThe corresponding author Katja Hanack is owner and manager of new/era/mabs. The other authors declare no competing financial or non-financial interests.AbbreviationCFSEcarboxyfluorescein succinimidyl esterDCdendritic cellELISAenzyme-linked immunosorbent assayFCSfetal calf serumHRPhorse raddish peroxidaseIgimmunoglobulinILinterleukinLPSlipopolysaccharideNCSneonatal calf serumRTroom temperatureVP1hamster polyomavirus capsid protein ER -