Membrane proteins

Our latest TechNote is here!

 
"Quick Characterization of Binding onto Unpurified GPCRs"
 

In this TechNote, we show how the WAVEsystem can be used for the kinetic analysis of G-protein binding onto detergent-solubilized, unpurified GPCRs. By shortening a typical 10-step purification process, this method requires significantly less material (30mL of cell culture) and less time (less than 1 hour), thereby offering an alternative for screening purposes at an early stage of the drug discovery process involving membrane proteins.

   

Membrane proteins are prime drug targets due to their essential role in regulating physiological processes. Familiar examples include G-protein coupled receptors (GPCR), histidine and tyrosine kinases, and cytokine receptors, in addition to a vast range of ion channels, enzymes, and transporters.

With more than half of all currently validated drugs on the market targeting membrane proteins, the importance of this protein family to drug development is well established. However, membrane proteins are notoriously difficult to study, mainly because their hydrophobicity makes them extremely unstable following their extraction from the cell membrane.

Understanding the interaction between a membrane protein and a small molecule drug or monoclonal antibody therapeutic is fundamental to drug development since it provides valuable insight regarding drug potency and efficacy. Yet to fully understand such liaisons, it is vital that the measurement of binding kinetics between these molecules is reliable.

TRANSFORMING DRUG DISCOVERY WITH THE CREOPTIX™ WAVEsystem

With the sensitivity and versatility to measure analyte-membrane protein interactions in a wide variety of sample matrices, the Creoptix™ WAVEsystem enables more detailed investigation of membrane protein pharmacology, providing, for example, real-time drug binding affinities and label-free kinetics. The Creoptix™ WAVEsystem also facilitates the discovery of new membrane proteins as therapeutic targets.


 

DETERGENTS AND OTHER ADDITIVES CAN COMPROMISE MEMBRANE PROTEIN INTEGRITY

The study of analyte-membrane protein interactions often involves the use of detergents, lipids and other additives to solubilize and purify the protein of interest. These non-native conditions have significant potential to alter the structure and conformation of the membrane protein, as well as impacting analyte-membrane protein binding kinetics, leading to inaccurate interpretation of data.

RETAIN STRUCTURE, CONFORMATION, AND ACTIVITY WITH THE CREOPTIX™ WAVEsystem

More than buffer
Explore a greater range of solubilization and purification conditions with our patented no-clog microfluidics, accommodating detergents, less common solvents, and multiple additives

Limited target consumption
Just 10-20 µg membrane protein is required to achieve a high protein density on the surface

High sensitivity
Push the limits and generate high-quality binding kinetics with our sensitive GCI technology and resolve data at very low responses

All sizes welcome
Screen, rank and characterize small analytes and fragments, even weak binders with fast dissociation rates – measure off-rates as fast as 10 sec-1 (half-life of 69 ms) –

Retain native conformation and activity.
Generate more physiologically relevant results

Save time
Study membrane protein pharmacology using only partially solubilized, unpurified material thanks to our no-clog microfluidics


 

VIRUS-LIKE PARTICLES, LIPOSOMES, AND NANODISCS PRESENT UNIQUE CHALLENGES

To avoid the lengthy solubilization and purification processes required to study membrane proteins, researchers often choose to incorporate these biomolecules into a membrane-like environment. This is typically provided by virus-like particles (VLPs), liposomes, or nanodiscs, all of which function to preserve membrane protein integrity and activity. A limitation of this approach is that the resultant size of these structures, combined with a tendency to aggregate, can cause microfluidics channels to clog. Impeded flow may lead to experimental delays and can also increase instrument maintenance costs, however the Creoptix® WAVEsystem overcomes this issue by incorporating a unique, no-clog microfluidic design.

ANALYZE LARGE MOLECULES AND BENEFIT FROM LOWER DETECTION LIMITS WITH THE CREOPTIX™ WAVEsystem

No clogging, regardless of size
Take advantage of the Creoptix™ WAVEsystem’s valveless microfluidics to analyze and characterize larger molecules.

High sensitivity
Explore lower limits of detection with our GCI technology and resolve kinetics at low responses.

 


 

STUDYING INTERACTIONS WITHIN A MEMBRANE CONTEXT PROVIDES GREATER INSIGHT

While some membrane proteins retain their analyte binding activity following solubilization and purification, this is not always the case. It is therefore important to study analyte-membrane protein binding within a membrane context to generate meaningful data. The Creoptix™ WAVEsystem allows researchers to investigate interactions between large binding partners and membrane proteins while maintaining the latter in their cell membrane environment. This provides essential insights to the binding of antibodies, nanobodies, or sybodies with the membrane protein of interest, and also allows investigation of membrane protein binding to intracellular signaling partners.

Close to native conditions
Immobilize membrane proteins from crude extracts with our no-clog microfluidic technology, without compromising the native state.

Wide range of affinities
Characterize large binding partners and resolve a wide range of affinities and off-rates with our stable GCI technology.
Learn more about sybodies and their kinetic properties

More interactions
Study the interaction between membrane proteins and their intracellular partners in a membrane context to better understand signaling pathways.