TY  - JOUR
A1  - Engel, Robert
A1  - Micheel, Burkhard
A1  - Hanack, Katja
T1  - Three-dimensional cell culture approach for in vitro immunization and the production of monoclonal antibodies
JF  - Biomedical materials : materials for tissue engineering and regenerative medicine
N2  - 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.
KW  - monoclonal antibody
KW  - hybridoma technology
KW  - in vitro immunization
KW  - 3D
KW  - cell culture
KW  - Geltrex
Y1  - 2022
U6  - https://doi.org/10.1088/1748-605X/ac7b00
SN  - 1748-6041
SN  - 1748-605X
VL  - 17
IS  - 5
PB  - Inst. of Physics
CY  - London
ER  -