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Tag-free, specific conjugation of glycosylated IgG1 antibodies using microbial transglutaminase

Hadjabdelhafid-Parisien, Adem ; Bitsch, Sebastian ; Macarrón Palacios, Arturo ; Deweid, Lukas ; Kolmar, Harald ; Pelletier, Joelle N. (2024)
Tag-free, specific conjugation of glycosylated IgG1 antibodies using microbial transglutaminase.
In: RSC Advances, 2022, 12 (52)
doi: 10.26083/tuprints-00027672
Artikel, Zweitveröffentlichung, Verlagsversion

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Kurzbeschreibung (Abstract)

We present an efficient approach for tag-free, site-specific conjugation of a fully glycosylated antibody using microbial transglutaminase (mTG). We created variants of trastuzumab where a single surface-exposed residue of the human crystallizable fragment had been substituted to glutamine, with the objective of enabling site-specific mTG-mediated conjugation with primary amine payloads. MTG reactivity was determined by conjugation to an amino fluorophore, demonstrating effective tag-free conjugation at the newly introduced I253Q site. The conjugation of one payload per antibody heavy chain was confirmed by mass spectrometry. We further demonstrated two-step mTG/click chemistry-based conjugation of I253Q trastuzumab with monomethyl auristatin E. Cytotoxicity and specificity of the resulting antibody–drug conjugate were indistinguishable from trastuzumab conjugated by another method although binding to the neonatal Fc receptor was impaired. The resulting fully glycosylated ADC is unique in that it results from minimal modification of the antibody sequence and offers potential for application to cellular imaging, fluorescence microscopy, western blotting or ELISA.

Typ des Eintrags: Artikel
Erschienen: 2024
Autor(en): Hadjabdelhafid-Parisien, Adem ; Bitsch, Sebastian ; Macarrón Palacios, Arturo ; Deweid, Lukas ; Kolmar, Harald ; Pelletier, Joelle N.
Art des Eintrags: Zweitveröffentlichung
Titel: Tag-free, specific conjugation of glycosylated IgG1 antibodies using microbial transglutaminase
Sprache: Englisch
Publikationsjahr: 20 August 2024
Ort: Darmstadt
Publikationsdatum der Erstveröffentlichung: 2022
Ort der Erstveröffentlichung: London
Verlag: Royal Society of Chemistry
Titel der Zeitschrift, Zeitung oder Schriftenreihe: RSC Advances
Jahrgang/Volume einer Zeitschrift: 12
(Heft-)Nummer: 52
DOI: 10.26083/tuprints-00027672
URL / URN: https://tuprints.ulb.tu-darmstadt.de/27672
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Herkunft: Zweitveröffentlichungsservice
Kurzbeschreibung (Abstract):

We present an efficient approach for tag-free, site-specific conjugation of a fully glycosylated antibody using microbial transglutaminase (mTG). We created variants of trastuzumab where a single surface-exposed residue of the human crystallizable fragment had been substituted to glutamine, with the objective of enabling site-specific mTG-mediated conjugation with primary amine payloads. MTG reactivity was determined by conjugation to an amino fluorophore, demonstrating effective tag-free conjugation at the newly introduced I253Q site. The conjugation of one payload per antibody heavy chain was confirmed by mass spectrometry. We further demonstrated two-step mTG/click chemistry-based conjugation of I253Q trastuzumab with monomethyl auristatin E. Cytotoxicity and specificity of the resulting antibody–drug conjugate were indistinguishable from trastuzumab conjugated by another method although binding to the neonatal Fc receptor was impaired. The resulting fully glycosylated ADC is unique in that it results from minimal modification of the antibody sequence and offers potential for application to cellular imaging, fluorescence microscopy, western blotting or ELISA.

Status: Verlagsversion
URN: urn:nbn:de:tuda-tuprints-276725
Sachgruppe der Dewey Dezimalklassifikatin (DDC): 500 Naturwissenschaften und Mathematik > 540 Chemie
500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie
Fachbereich(e)/-gebiet(e): Interdisziplinäre Forschungsprojekte
Interdisziplinäre Forschungsprojekte > Centre for Synthetic Biology
07 Fachbereich Chemie
07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie
07 Fachbereich Chemie > Clemens-Schöpf-Institut
Hinterlegungsdatum: 20 Aug 2024 13:33
Letzte Änderung: 21 Aug 2024 05:23
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