Scattering Methods Based Characterization of Liquid-Liquid Phase Separation
Recent developments in cell biology have drawn attention to the physical process of liquid-liquid phase separation (LLPS) as a relevant driver for non-chemical measurements of molecular assembly and cell compartmentalization. Characterization of biomolecular LLPS is essential for understanding biological processes. The proper characterization and control of aggregation-prone proteins requires a comprehensive understanding of their structural and transport properties, mainly their shape, conformation, degree of hydration, charge, diffusion coefficient and degree of aggregation in relation to protein volume concentration, electrolyte composition (ionic strength), pH and temperature. Scattering methods can be effectively used to determine these parameters in dilute aqueous solutions of proteins.
Customized Services
Our expert team is interested in the molecular characterization of the proteins that drive LLPS, yet direct detection of the dense phase of biomolecules is often technically difficult or impossible. By exploiting the symmetry between the conformational behavior of biomolecules in dilute and dense phases, we can infer details critical to phase separation by accurately measuring the dilute phase, thus avoiding the complex characterization of the dense phase.
Here, CD BioSciences offers scattering methods of different wavelengths of electromagnetic radiation for the characterization of biomolecular condensates in LLPS. Attractive to our customers, our light scattering methods are non-destructive, allowing samples to be used for additional experiments, including:
- Determination of protein structure.
- Determination of protein-protein interactions.
- Phase diagram of RNA phase separator formation.
- Characterization of the correlation distance within the phase isolate.
- Quantification of changes in protein and nucleic acid chain compression.
Dynamic Light Scattering (DLS) Platform
We offer the DLS method to detect the formation of biomolecular condensates, and to determine the saturation concentration of phase-separated proteins. In addition, we can analyze the reversibility of LLPS in monoclonal antibody solutions by comparing DLS plots under monodisperse conditions and after dilution of the layered sample into a monophasic, monodisperse, concentrated state.
Static Light Scattering (SLS) Platform
We offer a combination of DLS and SLS to characterize the double-nodal and spin-nodal profiles of phase separation systems, especially for elastin-like protein phase separation systems. Here, the inverse of the measured scattering intensity is plotted as a function of temperature, and the spin-nodal line temperature is determined by extrapolating to the infinite scattering intensity.
Small-Angle Scattering (SAS) Platform
Our SAS method is well suited for determining the size and shape of low-resolution (1-2 nm) biomolecular condensates, and provides useful insights into biomolecular interaction mechanisms and conformational transitions in terms of stoichiometry and alignment.
Depending on the merits of the different methods, we will design customized scattering-based solutions for your characterization of liquid-liquid phase separation. If you have any special requirements for our services, please feel free to contact us. We are looking forward to working together with your attractive projects.
For research use only, not intended for any clinical use.
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