Taylor-dispersion induced phase separation for efficient characterisation of protein condensate formation

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Taylor-dispersion induced phase separation for efficient characterisation of protein condensate formation

Angewandte Chemie Int. Ed. 2024
Rasmus Krogh Norrild, Thomas Oliver Mason, Lars Boyens-Thiele, Soumik Ray, Joachim Berg Mortensen, Anatol W. Fritsch, Juan M. Iglesias-Artola, Louise Kjær Klausen, Emil Georg Palmkvist Stender, Henrik Jensen, Alexander Kai Buell

This study introduces Taylor dispersion-induced phase separation (TDIPS), a method built on the commercially available Fida 1 instrument to characterise how proteins form biomolecular condensates via liquid-liquid phase separation. Flow Induced Dispersion Analysis (FIDA), based on Taylor dispersion, was used to measure condensate formation by exploiting the differing diffusivities of protein and buffer salts: as a nanoliter sample plug disperses in laminar capillary flow, salt separates from the slower protein and triggers phase separation, detected label-free through intrinsic fluorescence. The authors screened the phase behaviour of PGL-3, Ddx4N1, alpha-synuclein and lysozyme, mapped approximate phase diagrams, uncovered an unexpected re-entrant transition at very low ionic strength, and quantified LLPS-modifying compounds. TDIPS needs only nanoliters of unlabelled protein, enabling efficient, reproducible screening of condensation and its inhibitors.

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Taylor-dispersion induced phase separation for efficient characterisation of protein condensate formation
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