Hydrodynamic Radius Rh

General Biophysical Characteristics

Hydrodynamic Radius (Rh)

nm

Dynamic Light Scattering

20 mM HEPES, 10 mM NaCl

25 °C

8.0

1, 5, 8, 12, 16 and 20 mg/mL

None

None

The hydrodynamic radius (Rh) was determined by measuring Rh at a range of protein concentrations and then extrapolating the Rh value to zero protein concentration to avoid the effects of mAb self-association or repulsion on Rh determination.

Experiments were conducted using dynamic light scattering using a DynaPro plate reader (Wyatt Technology). Twenty µl of protein samples of 1, 5, 8, 12, 16 and 20 mg/mL in 20 mM HEPES, 10 mM NaCl, pH 8, were measured in 384-well clear-bottom plates (Corning) and the hydrodynamic radius was determined by linear extrapolating to 0 mg/mL. Ten acquisitions were recorded for 5 seconds at 25 °C with 0.5 and 1000 nm as low and high cut off values, respectively. Data processing was performed using Dynamics (version: 7.5.0.17; Wyatt Technology).