Production of a Chimeric Mouse–Fish Monoclonal Antibody by the CRISPR/Cas9 Technology

Ametrano, Alessia; Coscia, Maria Rosaria

doi:10.1007/978-1-0716-2313-8_19

Cited by 2 publications

(1 citation statement)

References 13 publications

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“…The Gibson product was transformed into DH5 α- competent E. coli . DNA plasmid was extracted from positive colonies by EndoFree Plasmid Maxi kit (QIAGEN) and double-digested with Hind III (NEB, #R0104S) and Sph I (NEB, #R0182S) at 37 °C for 1 h (Ametrano & Coscia, 2022).…”

Section: Methodsmentioning

confidence: 99%

Investigations on Antarctic fish IgM drives the generation of an engineered mAb by CRISPR/Cas9

Ametrano,

Miranda,

Moretta

et al. 2023

Preprint

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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 the one 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.

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Section: Methodsmentioning

confidence: 99%

Investigations on Antarctic fish IgM drives the generation of an engineered mAb by CRISPR/Cas9

Ametrano,

Miranda,

Moretta

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

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

Ametrano,

Miranda,

Moretta

et al. 2024

Front. Bioeng. Biotechnol.

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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.

show abstract

Production of a Chimeric Mouse–Fish Monoclonal Antibody by the CRISPR/Cas9 Technology

Cited by 2 publications

References 13 publications

Investigations on Antarctic fish IgM drives the generation of an engineered mAb by CRISPR/Cas9

Investigations on Antarctic fish IgM drives the generation of an engineered mAb by CRISPR/Cas9

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

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