Dynamics v7

To determine the liposome size and homogeneity, dynamic light scattering was performed on liposomes at 2.5 mg/mL at 20°C using a DynaPro NanoStar (Wyatt Technology). Light scattering data were analyzed using Dynamics v7.1.8 (Wyatt Technology).

To determine the liposome size and homogeneity, dynamic light scattering was performed on liposomes at 2.0 mg/mL at 20°C using DynaPro NanoStar (Wyatt Technology). Light scattering data were analyzed using Dynamics v7.1.8 (Wyatt Technology).

Dynamic light scattering was performed on liposomes at 2.0 mg/mL at
20°C using DynaPro NanoStar (Wyatt Technology) to determine the liposome
size and homogeneity. Light scattering data were analyzed using Dynamics v7.1.8
(Wyatt Technology).

Purified protein was measured using a DynaPro NanoStar (Wyatt) dynamic light scattering (DLS) setup. 0.5 mg/mL of I3-01 and 1wa3 were measured at 25 °C, then the temperature was ramped up to 90 °C, then ramped back down to 25 °C for temperature scans at 2 °C/minute. Measurements were taken in the presence of Tris-buffered saline (TBS): 25mM Tris, 500mM NaCl; buffered guanidinium hydrochloride (GuHCl): 25mM Tris, 500mM NaCl, 1–6.7 M GuHCl; or buffered guanidine thiocyanate (GITC): 25mM Tris, 500mM NaCl, 1–4 M GITC. Different concentrations of GITC equilibrated samples were achieved by combining stocks of 0 M and 4 M equilibrated solutions in different ratios while GuHCl equilibrated samples were equilibrated individually. Each sample was allowed to equilibrate in their respective buffer for at least 24 hours before measurement. Re-annealing experiments were performed by diluting I3-01 equilibrated in 3 M GITC down to 1 M GITC final concentration (0.166 mg/mL protein). Data analysis was performed using DYNAMICS v7 (Wyatt), reporting regularization fits (with D10/D50/D90) except for temperature ramp experiments, where cumulant fits were used. The ~1 nm radius particle consistent with GITC buffer alone was disregarded for analysis, and monodispersity was assumed when peak polydispersity was below 15%^{31 ,32} .

The hydrodynamic radius (R_h) was measured using a DynaPro plate reader (Wyatt Technologies, Santa Barbara, CA). Briefly, 60 μL of samples at 25 μM concentration was added in triplicate to a 384-well clear-bottom plate (Greiner Bio One, Monroe, NC). The wells were capped with 15 μL of mineral oil to prevent evaporation during long incubations. Data were analyzed using Dynamics V7 (Wyatt Technologies, Santa Barbara, CA).