Microscale thermophoresis as a powerful growing analytical technique for the investigation of biomolecular interaction and the determination of binding parameters

Abstract

The in vitro panel of technologies to address biomolecular interactions are in play, however microscale thermophoresis is continuously increasing in use to represent a key player in this arena. This review highlights the usefulness of microscale thermophoresis in the determination of molecular and biomolecular affinity interactions. This work reviews the literature from January 2016 to January 2022 about microscale thermophoresis. It gives a summarized overview about both the state-of the art and the development in the field of microscale thermophoresis. The principle of microscale thermophoresis is also described supported with self-created illustrations. Moreover, some recent advances are mentioned that showing application of the technique in investigating biomolecular interactions in different fields. Finally, advantages as well as drawbacks of the technique in comparison with other competing techniques are summarized.

Selection of spore-specific aptamers for Geobacillus stearothermophilus, a food spoilage bacterium

“…Several biophysical instruments or techniques could be investigated to measure aptamer-target binding affinity [61,62]. For instance, Microscale Thermophoresis (MST) [63,64] and Quartz Crystal Microbalance (QCM) [65,66] are two methods that could allow us to measure dissociation constants (Kd) for our aptamers towards Gbs spores.…”

Solution composition dependent Soret coefficient using commercial MicroScale Thermophoresis instrument

“…For instance, it is used to separate polymers and particles in eld-ow fractionation, [7][8][9] in combination with microuidics for submicron particle sorting, [10][11][12] and to monitor the protein-ligand binding in the pharmaceutical industry. [13][14][15] A deeper understanding of the thermophoretic dri of colloidal particles could be used to manipulate particle migration, such as trapping DNA and vesicles for their diagnostic potential.…”

Sensitive microscale thermophoresis assay for rapid ochratoxin A detection with fluorescently labeled engineered aptamer

“…10,11 MST has many applications in analyzing diverse molecular interactions, and MST and related techniques also provide a novel tool in the rapid quantification of targets, which usually requires an affinity ligand for specific recognition of targets. 11–19…”

“…10,11 MST has many applications in analyzing diverse molecular interactions, and MST and related techniques also provide a novel tool in the rapid quantification of targets, which usually requires an affinity ligand for specific recognition of targets. [11][12][13][14][15][16][17][18][19] Aptamers are single-stranded nucleic acids that can be selected from nucleic acid libraries using an in vitro process known as systematic evolution of ligands by exponential enrichment (SELEX), 20,21 and possess high affinity and specificity toward a broad range of targets from small molecules to proteins. [22][23][24] Due to the unique properties of aptamers, including easy chemical synthesis, engineerability, facile modification with chemical groups (e.g., fluorescent dyes), and high stability, they have been widely adopted in the construction of biosensors.…”