A protein is a very complex molecule. Therefore, several methods have to be used to fully characterize it. Methods concerning amino acid composition, glycosylation or other post-translational modifications are described in the MS section. However, intact mass, aggregates, fragments, net charge, identity and isoforms can also be detected by other methods:
– SDS-PAGE separates proteins with respect to their mass. Aggregates, fragments, and impurities are visible. With respect to a reference or a calibrator, identity and integrity can be assessed.
– Western Blotting can identify protein bands in an SDS-gel by transferring them to a membrane and staining with suitable binding partners.
– SEC is a chromatography method which separates proteins of different mass.
– Weak ion exchangers or IEF gels can separate differently charged proteins.
– Affinity to possible binding partners (if any), by DRX2 or Octet technology.
The biochemical and biophysical protein characterization is complimented by protein- and target specific functionality testing (e.g., target binding, signaling, cell-based assays). For more information see here.
At FyoniBio, we have long-term experiences in the characterization of proteins with a broad spectrum of analytical methods. Contact us to discuss the specific needs of your project and how we can support you.