Functional diversification of hybridoma produced antibodies by CRISPR/HDR genomic engineering
| Autor: | Duco van Dalen, Camille M. Le Gall, Tomislav Čaval, Johan M.S. van der Schoot, Anouk M. D. Becker, Michael Valente, J. Armando C. van Bruggen, Ferenc A. Scheeren, Carl G. Figdor, Marieke F. Fransen, Yusuf Dolen, Martijn Verdoes, Iris M. Hagemans, Maaike Nederend, Gestur Vidarsson, Jeanette H. W. Leusen, Albert J. R. Heck, Felix L. Fennemann, Arthur E. H. Bentlage, Melanie Engelfriet, Alper Cevirgel |
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| Jazyk: | angličtina |
| Předmět: |
0303 health sciences
biology Immunoglobulin domain Computational biology Isotype law.invention 03 medical and health sciences 0302 clinical medicine Antibody Isotype law 030220 oncology & carcinogenesis biology.protein Recombinant DNA CRISPR Hybridoma technology Antibody Function (biology) 030304 developmental biology |
| Zdroj: | bioRxiv |
| DOI: | 10.1101/551382 |
| Popis: | Hybridoma technology is instrumental for the development of novel antibody therapeutics and diagnostics. Recent preclinical and clinical studies highlight the importance of antibody isotype for therapeutic efficacy. However, since the sequence encoding the constant domains is fixed, tuning antibody function in hybridomas has been restricted. Here, we demonstrate a versatile CRISPR/HDR platform to rapidly engineer the constant immunoglobulin domains to obtain recombinant hybridomas which secrete antibodies in the preferred format, species and isotype. Using this platform, we obtained recombinant hybridomas secreting Fab’ fragments, isotype switched chimeric antibodies, and Fc-silent mutants. These antibody products are stable, retain their antigen specificity, and display their intrinsic Fc-effector functions in vitro and in vivo. Furthermore, we can site-specifically attach cargo to these antibody products via chemo-enzymatic modification. We believe this versatile platform facilitates antibody engineering for the entire scientific community, empowering preclinical antibody research.One Sentence SummaryWe demonstrate a universal CRISPR/HDR based platform for rapid genetic engineering of hybridomas to obtain functionally diverse antibody isotype panels in the species and format of choice. |
| Databáze: | OpenAIRE |
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