@article{EngelMicheelHanack2022,
  author    = {Engel, Robert and Micheel, Burkhard and Hanack, Katja},
  title     = {Three-dimensional cell culture approach for in vitro immunization and the production of monoclonal antibodies},
  series = {Biomedical materials : materials for tissue engineering and regenerative medicine},
  volume    = {17},
  journal   = {Biomedical materials : materials for tissue engineering and regenerative medicine},
  number    = {5},
  publisher = {Inst. of Physics},
  address   = {London},
  issn      = {1748-6041},
  doi       = {10.1088/1748-605X/ac7b00},
  pages     = {11},
  year      = {2022},
  abstract  = {The generation of monoclonal antibodies using an in vitro immunization approach is a promising alternative to conventional hybridoma technology. As recently published, the in vitro approach enables an antigen-specific activation of B lymphocytes within 10-12 d followed by immortalization and subsequent selection of hybridomas. This in vitro process can be further improved by using a three-dimensional surrounding to stabilize the complex microenvironment required for a successful immune reaction. In this study, the suitability of Geltrex as a material for the generation of monoclonal antigen-specific antibodies by in vitro immunization was analyzed. We could show that dendritic cells, B cells, and T cells were able to travel through and interact inside of the matrix, leading to the antigen-specific activation of T and B cells. For cell recovery and subsequent hybridoma technique the suitability of dispase and Corning cell recovery solution (CRS) was compared. In our experiments, the use of dispase resulted in a severe alteration of cell surface receptor expression patterns and significantly higher cell death, while we could not detect an adverse effect of Corning CRS. Finally, an easy approach for high-density cell culture was established by printing an alginate ring inside a cell culture vessel. The ring was filled with Geltrex, cells, and medium to ensure a sufficient supply during cultivation. Using this approach, we were able to generate monoclonal hybridomas that produce antigen-specific antibodies against ovalbumin and the SARS-CoV-2 nucleocapsid protein.},
  language  = {en}
}