Fig. 6. Developability properties of engineered variants from selected lead AB-000224.

a, Mutations made to generate engineered variants depicted in the Fv region of the Fab of AB-000224 in complex with NPNA4 (gold ribbon, PDB ID 6WFY; Methods). The light-chain framework is shown in silver and the heavy-chain in pale gray. The CDRs for light and heavy chains are indicated, respectively, as CDR1 (light and dark blue), CDR2 (light and dark purple) and CDR3 (light and dark cyan). Mutation sites of engineered variants are labeled as stability violations (red), deamidation sites (green) and paired sibling sites (gray; Methods). b–h, Developability properties of engineered variants of AB-000224 as characterized by scores from DSF WSS (b), thermal hold (c), chemically induced unfolding (d), low-pH stability (aggregate low-pH hold) (e), SINS (f), stand-up monolayer affinity chromatography (Zenix column RT) (g) and relative solubility analysis (PEG recovery) (h); green shading tindicates preferred ranges. i, Aggregate score of assay panel results from b–h for each engineered variant, with mutations shown in comparison with parental AB-000224. j,k, Characterization of stable cell production pools generated from engineered variants compared with AB-000224 for production titers (g l⁻¹) (j) and cell-specific productivity (q_P, pg per cell per day) (k). Boxes indicate interquartile ranges, lines within boxes are medians and whiskers represent minimum and maximum (four independent replicates of each antibody).