Binding-based Ligand Screening Services
G-protein coupled receptors (GPCRs) are a broad class of protein receptors found in all parts of the body that are involved in most essential processes. More than thirty percent of medications are targeted at this family of receptors, demonstrating the significance of these targets in pharmaceutical development. GPCR ligand screening has gained more attention due to the potential to screen for biased agonists, which only activate the main signaling pathway and thereby minimize harmful side effects. Indeed, the creation of diverse assays has resulted from ligand screening for different classes of GPCRs and the introduction of novel ideas regarding GPCR activation. Creative Biolabs can offer a wide of binding-based assays for GPCR ligand screening.
Binding-based Ligand Screening Services in Creative Biolabs
- GPCR Ligand Screening Services Using SPR
One extremely sensitive technique for tracking binding between two species in real time is surface plasmon resonance, or SPR. The refractive index shift at a gold/glass/solvent interface, which modifies the angle at which light is lost to surface plasmons, is measured using surface plasmon resonance (SPR). Target proteins are typically fixed on sensors via affinity capture strategies such as polyhistidine-tags, biotin, or Fc receptors, or by covalently attaching the protein to lysine. The stability of proteins may place a limit on the hundreds of molecules that can be measured daily using this format. SPR is somewhat costly and necessitates tight control over temperature, particulate matter, and buffers despite its high sensitivity.
- GPCR Ligand Screening Services Using MST
MST is useful for measuring a wide range of biochemicals, primarily the movement of particles in response to tiny temperature fields. Proteins' ability to bind to tiny molecules and even tiny ions, like calcium, can be measured using MST. Protein-protein interactions, interactions between membrane proteins (including SNARE proteins), competitive inhibition, interactions between proteins and lipids in micelle- or short-chain- or relatively flat-layered (e.g., nanodisc) bilayers, and in certain cases, even kinetic or thermodynamic parameters can all be measured using MST.
- GPCR Ligand Screening Services Using TR-FRET
Through dipole-dipole interactions, energy from a fluorescent donor is absorbed by an acceptor in fluorescence resonance energy transfer, or FRET. The efficiency of energy transfer is dependent on the spectrum overlap, distance, and relative orientation between the donor and acceptor. Common donor-acceptor pairs often have useful distances between them of 2-6 nm, which are suitable for a variety of protein-protein interactions. Usually, dyes are chemical compounds, such as rhodamine or fluorescein, conjugated to the target protein pair or fluorescent proteins fused to the target protein.
- GPCR Ligand Screening Services Using ALIS
The Automated Ligand Identification System (ALIS) is a technology based on AS-MS that combines high-resolution mass spectrometric detection with online two-dimensional chromatography. Target-based drug discovery faces several problems, but ALIS, a completely automated procedure based on full-scan high-resolution-accurate mass spectrometry, has been employed to overcome them. ALIS is ideally positioned to quickly screen huge collections of chemicals to identify ligands to numerous proteins in parallel, such as a clinically interesting pathway, because of its high throughput and flexible technique development.
- GPCR Ligand Screening Services Using MS
A common method for looking for interactions between proteins and tiny molecules is mass spectrometry (MS). A popular setup is affinity selection-mass spectrometry (AS-MS), which involves incubating a protein along with a pool of chemicals. Then, protein-ligand complexes are quickly isolated from unbound compounds, and bound ligands are detected by dissociating them using LC-ESI-MS. There are several techniques for separating protein-ligand complexes from unbound ligands, including size-exclusion chromatography and (pulsed) ultrafiltration. In favorable circumstances, mass spectrometry may also quantify protein-molecule adducts. By using MS, binding is confirmed and covalent ligand-protein complexes are recognized.
Independent of their impact on the function of the protein, binding-based tests examine how the library components bind to the target protein. We may provide ligand screening services utilizing surface plasmon resonance, nuclear magnetic resonance (NMR), and X-ray crystallography in addition to the techniques mentioned above. Please contact us for more information. Our scientist team is able to quickly and effectively design and create the best method for your project.
