Reliable repurposing of the antibody interactome inside the cell

O'Shea, Caitlin M and Shahzad, Rushba and Aghasoleimani, Kimia and Newman, Stuart and Panmanee, Jiraporn and Schalkwyk, Leonard C and Brooke, Greg N and Benson, Fiona E and Trimmer, James S and Bosco, Daryl A and Fujisawa, Takao and Ichijo, Hidenori and Cashman, Neil R and Engel, Stanislav and Wright, Gareth S A (2026) Reliable repurposing of the antibody interactome inside the cell. Nature Communications. ISSN 2041-1723

Abstract

Eighty-five percent of the human proteome has at least one interacting monoclonal antibody. These molecules penetrate the cytoplasm poorly and are very often non-functional within the cell. Analysis of antibody variable domains and characterisation of forty-five single-chain variable fragment (scFv) intrabodies expressed in human cells indicated charge to have the greatest impact on solubility. We created new interdomain linkers, optimised scFv domain orientation and found an optimisable charge discrepancy between variable heavy framework and CDR sites. When applied to reduce the search space and rank the products of AI-led inverse folding this creates a single highly soluble, abundant and stable intrabody with parent antibody epitope recognition. Over six hundred intrabody sequences are presented targeting sixty cytoplasmic proteins with linear, conformational, post-translational modification or oligomer specificity. Interactions were validated for p53, α-synuclein, SOD1, polyQ, FUS/TLS, UCHL1 and GFP. Here we show reliable repurposing of the sequenced antibody interactome inside the cell.