Technology

Membrane proteins and especially so called G-Protein Coupled Receptors (GPCRs) represent the most important drug targets. Despite their importance, they belong still to the most challenging drug targets.

InSingulo proprietary single molecule surface-sensitive microscopy-based method contributes to this challenge by probing the interaction between potential drug compounds and membrane proteins with single-molecule resolution1. It is enabling inhibition in solution perturbations of the interaction to reveal detailed kinetic information, coined dynamic inhibition in solution assay, or dISA in short. In essence, this provides label-free interaction analysis of drug-target interactions providing several advantages compared to conventional methods.

In essence, InSingulo provides label-free interaction analysis of drug-target interactions providing several advantages compared to conventional methods

- Tim Kaminski, Chief Scientific Officer

First advantage

First, instead of adopting membrane proteins to a membrane-free environment, spending tremendous efforts of building a surrogate environment or trying to stabilizing them to sustain in an artificial environment, In Singulo Solutions proprietary technology allows keeping membrane proteins throughout the whole experimental workflow in the right lipid-based environment: the native cell membrane.

Second advantage

Second, unlike in conventional label-free assays the liposomes can remain in solution and only a tool compound has to be immobilized at the sensor surface. The interaction between the membrane protein and tool compound can be challenged with ligands of choice and unlike with other inhibition in solution assays, our technology allows to measure the interaction kinetics between the ligand and the target totally independent from the properties of the tool-compound properties. In combination with the fact that non-specific interaction between potential drug compounds and the lipid membrane surrounding or off target binding does not influence the measurements, this means that the method offer higher specificity than conventional label free methods even in complex environments.

Third advantage

Third, the single-molecule sensitivity, makes possible for In Singulo Solutions’ technology to operate using tool-compounds within a broad affinity range from sub-nanomolar to micromolar, and the same tool compound can be used throughout the whole development process from screening to end of lead optimization.

Fourth advantage

Forth, being able to measure at the single molecule level our technologies needs only small sample amounts and low target protein concentration. This enables us to overcome limitations like the tight-binding problem or ligand depletion, too.

Summary

In summary, by eliminating the need of isolation and purification of membrane proteins, one of the most common pitfalls in working with membrane proteins is abolished. Further, by delivering full kinetic interaction analysis with the highest possible sensitivity, the interaction of single molecules, our technology allows to accelerate drug development projects in an unprecedented way. Finally, being developed in close collaboration with leading drug developers our technology is made to overcome key challenges scientist in drug discovery experience when working with membrane proteins.

Applications

How our technology is put to use

Membrane proteins

Membrane proteins represent the majority of drug targets. We have developed a process that simplifies the development process and increases the chances for success.

Intrinsically Disordered Proteins

Intrinsically disordered proteins or proteins with intrinsically disordered domains are highly abundant. Using our dISA technology allows to determine the binding kinetics of various ligands straightforward.