A novel selection strategy for antibody producing hybridoma cells based on a new transgenic fusion cell line
- The use of monoclonal antibodies is ubiquitous in science and biomedicine but the generation and validation process of antibodies is nevertheless complicated and time-consuming. To address these issues we developed a novel selective technology based on an artificial cell surface construct by which secreted antibodies were connected to the corresponding hybridoma cell when they possess the desired antigen-specificity. Further the system enables the selection of desired isotypes and the screening for potential cross-reactivities in the same context. For the design of the construct we combined the transmembrane domain of the EGF-receptor with a hemagglutinin epitope and a biotin acceptor peptide and performed a transposon-mediated transfection of myeloma cell lines. The stably transfected myeloma cell line was used for the generation of hybridoma cells and an antigen- and isotype-specific screening method was established. The system has been validated for globular protein antigens as well as for haptens and enables a fast and early stageThe use of monoclonal antibodies is ubiquitous in science and biomedicine but the generation and validation process of antibodies is nevertheless complicated and time-consuming. To address these issues we developed a novel selective technology based on an artificial cell surface construct by which secreted antibodies were connected to the corresponding hybridoma cell when they possess the desired antigen-specificity. Further the system enables the selection of desired isotypes and the screening for potential cross-reactivities in the same context. For the design of the construct we combined the transmembrane domain of the EGF-receptor with a hemagglutinin epitope and a biotin acceptor peptide and performed a transposon-mediated transfection of myeloma cell lines. The stably transfected myeloma cell line was used for the generation of hybridoma cells and an antigen- and isotype-specific screening method was established. The system has been validated for globular protein antigens as well as for haptens and enables a fast and early stage selection and validation of monoclonal antibodies in one step.…
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| Author details: | Martin Listek, Anja Hönow, Manfred GossenORCiDGND, Katja HanackORCiDGND |
|---|---|
| URN: | urn:nbn:de:kobv:517-opus4-459893 |
| DOI: | https://doi.org/10.25932/publishup-45989 |
| ISSN: | 1866-8372 |
| Title of parent work (German): | Postprints der Universität Potsdam : Mathematisch Naturwissenschaftliche Reihe |
| Publication series (Volume number): | Zweitveröffentlichungen der Universität Potsdam : Mathematisch-Naturwissenschaftliche Reihe (865) |
| Publication type: | Postprint |
| Language: | English |
| Date of first publication: | 2020/04/07 |
| Publication year: | 2020 |
| Publishing institution: | Universität Potsdam |
| Release date: | 2020/04/07 |
| Tag: | Antibody generation; Assay systems |
| Issue: | 865 |
| Number of pages: | 14 |
| Source: | Scientific Reports 10 (2020) 1664 DOI:10.1038/s41598-020-58571-w |
| Organizational units: | Mathematisch-Naturwissenschaftliche Fakultät |
| DDC classification: | 5 Naturwissenschaften und Mathematik / 50 Naturwissenschaften / 500 Naturwissenschaften und Mathematik |
| 6 Technik, Medizin, angewandte Wissenschaften / 60 Technik / 600 Technik, Technologie | |
| Peer review: | Referiert |
| Publishing method: | Open Access |
| License (German): |  CC-BY - Namensnennung 4.0 International |
| External remark: | Bibliographieeintrag der Originalveröffentlichung/Quelle |
