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Three-dimensional cell culture approach for in vitro immunization and the production of monoclonal antibodies
- 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 ofThe 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.…
Author details: | Robert EngelORCiD, Burkhard Micheel, Katja HanackORCiDGND |
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DOI: | https://doi.org/10.1088/1748-605X/ac7b00 |
ISSN: | 1748-6041 |
ISSN: | 1748-605X |
Pubmed ID: | https://pubmed.ncbi.nlm.nih.gov/35728755 |
Title of parent work (English): | Biomedical materials : materials for tissue engineering and regenerative medicine |
Publisher: | Inst. of Physics |
Place of publishing: | London |
Publication type: | Article |
Language: | English |
Date of first publication: | 2022/07/04 |
Publication year: | 2022 |
Release date: | 2023/01/25 |
Tag: | 3D; Geltrex; cell culture; hybridoma technology; in vitro immunization; monoclonal antibody |
Volume: | 17 |
Issue: | 5 |
Article number: | 055003 |
Number of pages: | 11 |
Funding institution: | Helmholtz Graduate School for Macromolecular Bioscience [VH-GS-503] |
Organizational units: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie und Biologie |
DDC classification: | 6 Technik, Medizin, angewandte Wissenschaften / 61 Medizin und Gesundheit / 610 Medizin und Gesundheit |
Peer review: | Referiert |
Publishing method: | Open Access / Hybrid Open-Access |
License (German): | CC-BY - Namensnennung 4.0 International |