Synthesis and application of PEGylated tracer particles for measuring protein solution viscosities using Dynamic Light Scattering-based microrheology.

The measurement of flow properties, such as the zero shear viscosity, of protein solutions is of paramount importance for many applications such as pharmaceutical formulations, where the syringeability of physiologically effective doses is a key property. However, the determination of these properties with classical rheological methods is often challenging due to e.g. detrimental surface effects or simply the lack of sufficient material. A possible alternative is Dynamic Light Scattering-based microrheology, where the Brownian motion of tracer particles embedded in the protein solution is monitored to access the zero shear viscosity of the sample. The prime advantages of this method compared to classical rheology are the absence of disturbing surface effects and the up to two orders of magnitude smaller protein quantities needed for an entire concentration series. This Protocol provides a detailed description of the synthesis of sterically stabilized tracer particles with surface and overall particle properties specifically designed to investigate the viscosity of protein solutions up to concentrations close to the arrest transition. These particles are tailored to avoid protein-particle as well as particle-particle aggregation at various sample conditions and thus allow for an artifact-free application of Dynamic Light Scattering-based tracer microrheology to determine the flow behaviour of biological samples. The Protocol concludes with step by step instructions for the characterization of protein solutions using a combination of the tracer particles and an advanced dynamic light scattering technique yielding the concentration-dependent zero shear viscosity.