In vitro Immunization - Generation of monoclonal antibodies

Monoclonal antibodies were conventionally generated by hybridoma technology. The process includes the immunization of host animals with the antigen of interest in order to initiate an appropriate and specific immune response. Normally, this covers a period of 3 month till the mammalian immune system has built up a highly specific immune response. To transfer this process into an in vitro approach would lead to a reduction in time and to a process which could be monitored better than in the host individual. Further an in vitro approach allows the usage of cells from alternative hosts as humans or camelids as well. Therefore, the establishment of human and camelid in vitro approaches is also part of our research.

In our in vitro approach all relevant immune cell populations from bone marrow and spleen were isolated, characterized and co-cultivated. The next step is an antigen-specific activation of B lymphocytes followed by an immortalization to hybridoma cells. In the current research project this approach is modified to a cell line based approach to have unlimited access to the important cell types. The single activation steps were characterized by immunotechnological methods such as flow cytometry, immunofluorescence, ELISA or immunoblotting. Intracellular markers such as transcription factors or cytokines were analyzed by RT-PCR or intracellular staining.

In combination with our VLP immunization and selection we are developing an all-round platform solution for the generation of monoclonal antibodies in a significant shorter time frame.