Generation of antigen-specific monoclonal antibodies by in vitro immunization (TECH2P.904)
| Autor: | Katja Heilmann, Pamela Holzlöhner |
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| Rok vydání: | 2015 |
| Předmět: | |
| Zdroj: | The Journal of Immunology. 194:206.14-206.14 |
| ISSN: | 1550-6606 0022-1767 |
| DOI: | 10.4049/jimmunol.194.supp.206.14 |
| Popis: | Monoclonal antibodies are one of the most important tools in biomedicine but the generation by hybridoma technology is very time consuming and laborious. The adaptation of the process to an in vitro approach is therefore a smart alternative to reduce the disadvantages of hybridoma technology. In this study a protocol for in vitro immunization was developed which enables hematopoietic stem cells to differentiate into naïve dendritic cells. These cells were specifically activated with the antigen of interest and cocultivated with naïve B and T lymphocytes in order to generate antigen specific B lymphocytes. After 4 days the cells were fused with myeloma cells and antigen specific hybridomas were selected. The produced antibody was purified and characterised in comparison to a conventional monoclonal antibody reacting against the same antigen. Further this protocol could be adapted to human cells and the generation of antigen specific hybridoma cells secreting human IgG molecules could be established. The developed protocols allowed a generation of antigen specific hybridoma cells of murine and human origin within 3 weeks. Therefore it seems that the immunization in vitro is useful for avoiding the disadvantages of conventional hybridoma technology. |
| Databáze: | OpenAIRE |
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