Experimental high throughput approaches to quantify the thermodynamics of biomolecular condensate formation [EXTRAORDINARYNodes Seminar]

Biomolecular condensates (BMCs), spatial regions that are not membrane-bound but nevertheless strongly enriched in certain biomolecular components, are of great current interest, due to their relevance in biological function and disease. Experimental methods that allow to study the dependence of BMC formation on solution conditions and amino acid sequence are therefore sought after, in particular if those methods allow to screen these parameters at high throughput (HTP). Such HTP methods are valuable for screening campaigns, but also to obtain improved understanding of the driving forces responsible for BMC formation.

In this seminar, I will present two microfluidics-based methods, Capflex (Stender et al., Nat. Comm. 2021) and TDIPS (Norrild et al., Angew. Chem. Int. Ed. 2024), that are based on the commercially available FIDA1 microcapillary based instrument and that allow the convenient study of the influence of solution conditions on BMC formation. Furthermore, I will present a novel method based on mRNA-display that we call Condensate Partitioning by mRNA-Display (CPmD, Norrild et al., bioRxiv pre-print 2024, https://doi.org/10.1101/2024.12.21.629870). This method allows to study the partitioning of tens of thousands of peptides into BMCs in a single experiment and provides fundamental insights into the basic thermodynamics of BMC formation.”