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 |
| Zugehörige Links: | |
| 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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| Suche nach Titel in: | TUfind oder in Google |
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- Tag-free, specific conjugation of glycosylated IgG1 antibodies using microbial transglutaminase. (deposited 20 Aug 2024 13:33) [Gegenwärtig angezeigt]
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