The kinetics, thermodynamics, and molecular mechanisms of liquid-liquid phase separation (LLPS) are critical in cell biology. We are developing reproducible stream-based methods to analyze the LLPS kinetics of proteins that are often severely aggregation-prone. Here at CD BioSciences, we offer a powerful and easy-to-use microfluidic experimental platform to enable such measurements.

Introduction

LLPS has been identified as a potential driver of cellular compartmentalization in membrane-free regions and plays a key role in cellular biology processes. Phase separation and its dysregulation may be associated with cancer, neurodegenerative diseases and aging. Understanding the impact of disease mutations and fusion events, and improving our understanding of the physiological processes involved in phase separation, requires characterizing the physicochemical rules that drive the interactions of LLPS. A wide range of microfluidic techniques have been applied to study macromolecular phase transition phenomena, from normally open to normally closed valves and emulsion droplet workflows for combined composition titrations. Alternatively, semi-permeable membrane transport by water vapor permeation or dialysis has been successfully used to study phase transitions.

Fig. 1. Schematic illustration of some key features of microfluidic systems when analyzing biomolecular condensates. (Linsenmeier M, et al., 2021)

Customized Services

In vitro reconstitution methods can mimic key biological features to enable analysis of fundamental aspects of drops. Based on cutting-edge microfluidics, CD BioSciences offers the more economical microfluidic approach to characterize LLPS. This approach allows our researchers to manipulate various parameters of the solution, such as protein/RNA concentration, temperature, pH, ionic strength and solution composition, for a wide range of studies of different aspects of LLPS, including:

✓ Phase diagram.

✓ Assembly and disassembly kinetics.

✓ Rheology and surface properties.

✓ Exchange of materials with their surroundings.

✓ Coupling between partitioning and biochemical reactions.

Our microfluidic tools are extremely attractive for the analysis of biomolecular phase changes. In addition to the ability to perform high-throughput measurements on small sample volumes, they also allow precise control of self-assembly in time and space, resulting in accurate quantitative analysis of phase transitions in biomolecular condensates. Our microfluidic approach also allows manipulation of samples at different stages of the biomolecular coalescence lifetime, enabling long time observation of the formed coalescence.

Advantages of the Microfluidic Experiment

• It is a valuable tool for the analysis of biological self-assembly and the fabrication of bottom-up synthetic compartments.
• Can easily perform experiments at high throughput and using limited amounts of material.
• Manipulation of liquid samples at the micron scale allows manipulation that is essentially impossible at the large volume scale.
• Superior manipulation of the protein environment is produced in both space and time.

Our microfluidic platform can help you gain insight into the phase separation process and the nature of the resulting compartments. This technology will be particularly important when analyzing phase transitions in biology and their role in cellular metabolism, as well as new therapeutic strategies for various diseases associated with phase separation. If you are interested in our services, please do not hesitate to contact us for more information.