Fig. 5: Plugged antibody nanoparticles exhibit tunable dissociation in response to acidification.
From: Computational design of non-porous pH-responsive antibody nanoparticles
a, O432-17 nanoparticles were assembled with AF647-conjugated trimeric plug variants, designed tetramer, sfGFP-Fc and α-Myc antibody. b, O432-17 nanoparticles were immobilized on Myc-peptide-coated beads and incubated in titrated pH buffers. Beads were washed by centrifugation and resuspended in 25 mM Tris and 500 mM NaCl, pH 8, and the remaining trimer and sfGFP-Fc fluorescence on the beads was analyzed by flow cytometry. c–g, AF647 and sfGFP fluorescence levels were normalized to the minimum and maximum values across the titration and analyzed as a function of the pH for each nanoparticle variant (O432-17(–)_2HIS (c), O432-17(–)_0HIS (d), O432-17(–)_3HIS_I57V (e), O432-17(–)_3HIS_L75A (f), O432-17(–)_3HIS_I57V_L75A (g)). Data are normalized to the minimum and maximum fluorescence per titration and presented as mean ± s.e.m. for three biologically independent replicates. UCSF ChimeraX 1.6 (ref. 54) and the PyMOL Molecular Graphics System version 2.5 (Schrödinger) were used to create a; b was created using BioRender; GraphPad Prism version 9.3.1 (GraphPad Software) was used to create c–g.
