Engineering of pH-dependent antigen binding properties for toxin-targeting IgG1 antibodies using light-chain shuffling

Tulika Tulika, Fulgencio Ruso-Julve, Shirin Ahmadi, Anne Ljungars, Esperanza Rivera-de-Torre, Jack Wade, Monica L. Fernández-Quintero, Timothy P. Jenkins, Selma B. Belfakir, Georgina M.S. Ross, Lars Boyens-Thiele, Alexander K. Buell, Siri A. Sakya, Christoffer V. Sørensen, Markus-Frederik Bohn, Line Ledsgaard, Bjørn G. Voldborg, Chiara Francavilla, Tilman Schlothauer, Bruno LomonteJan Terje Andersen*, Andreas H. Laustsen*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

Immunoglobulin G (IgG) antibodies that bind their cognate antigen in a pH-dependent manner (acid-switched antibodies) can release their bound antigen for degradation in the acidic environment of endosomes, while the IgGs are rescued by the neonatal Fc receptor (FcRn). Thus, such IgGs can neutralize multiple antigens over time and therefore be used at lower doses than their non-pH-responsive counterparts. Here, we show that light-chain shuffling combined with phage display technology can be used to discover IgG1 antibodies with increased pH-dependent antigen binding properties, using the snake venom toxins, myotoxin II and α-cobratoxin, as examples. We reveal differences in how the selected IgG1s engage their antigens and human FcRn and show how these differences translate into distinct cellular handling properties related to their pH-dependent antigen binding phenotypes and Fc-engineering for improved FcRn binding. Our study showcases the complexity of engineering pH-dependent antigen binding IgG1s and demonstrates the effects on cellular antibody-antigen recycling..
Original languageEnglish
JournalStructure
Volume32
Issue number9
Pages (from-to)1404-1418.e7
ISSN0969-2126
DOIs
Publication statusPublished - 2024

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