DataSheet1_A structural peculiarity of Antarctic fish IgM drives the generation of an engineered mAb by CRISPR/Cas9.docx

• Main Authors: Alessia Ametrano, Bruno Miranda, Rosalba Moretta, Principia Dardano, Luca De Stefano, Umberto Oreste, Maria Rosaria Coscia
• Format: Dataset
• Language: unknown
• Published: 2024
• Subjects: Biotechnology; Biological Engineering; Genetic Engineering; Biomarkers; Biomaterials; Biomechanical Engineering; Biomedical Engineering not elsewhere classified; Synthetic Biology; Agricultural Marine Biotechnology; Bioremediation; Bioprocessing; Bioproduction and Bioproducts; Industrial Biotechnology Diagnostics (incl. Biosensors); Industrial Microbiology (incl. Biofeedstocks); Industrial Molecular Engineering of Nucleic Acids and Proteins; Industrial Biotechnology not elsewhere classified; Medical Biotechnology Diagnostics (incl. Biosensors); Medical Molecular Engineering of Nucleic Acids and Proteins; Regenerative Medicine (incl. Stem Cells and Tissue Engineering); Medical Biotechnology not elsewhere classified; genome editing; IgH gene locus; hinge region; plasmonic substrate; optical biosensor; antigen-binding affinity; teleost fish; Antarc*; Antarctic
• Online Access: https://doi.org/10.3389/fbioe.2024.1315633.s001; https://figshare.com/articles/dataset/DataSheet1_A_structural_peculiarity_of_Antarctic_fish_IgM_drives_the_generation_of_an_engineered_mAb_by_CRISPR_Cas9_docx/26368012

IgM is the major circulating Ig isotype in teleost fish, showing in Antarctic fish unique features such as an extraordinary long hinge region, which plays a crucial role in antibody structure and function. In this work, we describe the replacement of the hinge region of a murine monoclonal antibody (mAb) with the peculiar hinge from Antarctic fish IgM. We use the CRISPR/Cas9 system as a powerful tool for generating the engineered mAb. Then, we assessed its functionality by using an innovative plasmonic substrate based on bimetallic nanoislands (AgAuNIs). The affinity constant of the modified mAb was 2.5-fold higher than that obtained from wild-type mAb against the specific antigen. Here, we show the suitability of the CRISPR/Cas9 method for modifying a precise region in immunoglobulin gene loci. The overall results could open a frontier in further structural modifications of mAbs for biomedical and diagnostic purposes.