Three-dimensional cell culture approach for in vitro immunization and the production of monoclonal antibodies. (1st September 2022)

• Record Type: Journal Article
• Authors: Engel, Robert; Micheel, Burkhard; Hanack, Katja
• Is Part Of: Biomedical materials. Volume 17:Number 5(2022)
• Publication Date: 2022-09-01
• Subjects: monoclonal antibody -- hybridoma technology -- in vitro immunization -- 3D cell culture -- Geltrex; Biomedical materials -- Periodicals; 610.28
• DOI: 10.1088/1748-605X/ac7b00
• Languages: English
• ISSNs: 1748-6041

Abstract

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.