Biomolecular Interactions Analytics using MicroScale Thermophoresis

Piotr Wardega

NanoTemper Technologies, Bobrzynskiego 14, Krakow, Poland

NanoTemper Technologies proprietary technology; Microscale Thermophoresis (MST) allows for quantitative analysis of biomolecular interactions in free solutions. The technique is based on phenomenon of thermophoresis, the directed motion of molecules in temperature gradient. Thermophoresis is highly sensitive to all types of binding-induced changes of molecular properties, be it in size, surface charge and hydration shell or conformation. Upon any type of interaction at least one of the above mentioned parameters will change. In an all optical approach, MST is induced using an infrared laser for local thermal gradient induction, and molecules mobility across it is analysed through fluorescence detection. In addition to fluorescence by various available fluorophores or fusion proteins attached to one of the binding partners, intrinsic protein fluorescence can be utilized for MST thus allowing for label-free MST analysis.

Our technique’s flexibility in assay design and experimental execution qualifies MST for biomolecular interaction analysis in complex experimental settings, which we herein demonstrate by addressing typically challenging types of binding events from various fields of life science. Furthermore, MST assays are highly adaptable to fit to the diverse requirements of different biomolecules, e.g. membrane proteins stabilized in solution, multi-component protein structures or nanoparticles. The type of buffer and additives can be chosen freely and does not discriminate its composition. Measuring is also possible in complex bioliquids like cell lysate, blood serum or even direct analysis without protein purification and thus under close to in vivo conditions. Binding modes that are quantifiable via MST include dimerization, cooperativity and competition as well. Thermophoresis makes both high- (pM) and low affinity (mM), interactions available in shape of single instrument solution. For more insight and details you are welcome to visit our booth.