Biomolecular condensate phase diagrams with a combinatorial microdroplet platform

The assembly of biomolecules into condensates is a fundamental process underlying the organisation of the intracellular space and the regulation of many cellular functions. Mapping and characterising phase behaviour of biomolecules is essential to understand the mechanisms of condensate assembly, and to develop therapeutic strategies targeting biomolecular condensate systems. A central concept for characterising phase-separating systems is the phase diagram. Phase diagrams are typically built from numerous individual measurements sampling different parts of the parameter space. However, even when performed in microwell plate format, this process is slow, low throughput and requires significant sample consumption. To address this challenge, we present here a combinatorial droplet microfluidic platform, termed PhaseScan, for rapid and high-resolution acquisition of multidimensional biomolecular phase diagrams. Using this platform, we characterise the phase behaviour of a wide range of systems under a variety of conditions and demonstrate that this approach allows the quantitative characterisation of the effect of small molecules on biomolecular phase transitions.

Automated summary

The assembly of biomolecules into condensates plays a crucial role in organizing the intracellular space and regulating cellular functions. Characterizing the phase behavior of biomolecules is essential for understanding condensate assembly mechanisms and developing therapeutic strategies. However, traditional methods for constructing phase diagrams are slow, low throughput, and sample-intensive. To address this challenge, a combinatorial droplet microfluidic platform called PhaseScan is introduced for rapid and high-resolution acquisition of multidimensional biomolecular phase diagrams.