Reinvestigation of the Automated Synthesis of Stoichiometrically Conjugated Antibodies to Access High Molecular Weight Payloads and Multiplexed Conjugation via an In-Solution Trans-Tagging Process.
Fiche publication
Date publication
octobre 2023
Journal
ACS omega
Auteurs
Membres identifiés du Cancéropôle Est :
Dr CIANFERANI Sarah, Dr WAGNER Alain, Dr KOLODYCH Sergii, Dr KONIEV Sasha, Dr CHAUBET Guilhem
Tous les auteurs :
Lehot V, Lidický O, Most J, Erb S, Dovgan I, Osypenko A, Koniev O, Kolodych S, Kotrchová L, Chaubet G, Cianférani S, Etrych T, Wagner A
Lien Pubmed
Résumé
Protein conjugates have found applications in a wide variety of fields, ranging from therapeutics to imaging and detection. However, robust control over the parameters of the conjugation process (such as sites and degree of conjugation) remains challenging. Previously, our group introduced Equimolar NAtive Chemical Tagging (ENACT), a method which allows for the monofunctionalization of proteins by combining an iterative low-conversion bioconjugation, an automated process, and a bioorthogonal trans-tagging reaction. However, while the automated ENACT was dimensioned to achieve monoconjugation at the mg scale, in early stage research, because of the rarity and cost of the starting materials, it is often necessary to prepare conjugates at the lower, μg, scale. Here, we introduce modified ENACT protocols, as well as a new ENACT conjugation reagent, which allow for the monofunctionalization of proteins on the micrograms scale, using minimal quantities of payload.
Référence
ACS Omega. 2023 10 31;8(43):40508-40516