Expression of natural and recombinant proteins is an important requisite for structural biology applications. More often than not, proteins are challenging to express outside their original host or in sufficient quantity even within their native host. The Structural Biology department’s Protein Technologies Center solves difficult protein expression problems by employing semi-automated, multi-pronged approaches at each stage of the pipeline, from construct design to cloning, expression, purification, and characterization of all protein targets, including membrane proteins, multi-protein complexes, and assemblies.
The Center is developing a large set of expression vectors for E. coli, insect cell, mammalian cell, and cell-free expression systems. For protein purification, the Center utilizes FPLC and HPLC systems with a combination of chromatographic methods such as affinity chromatography, ion exchange and hydrophobic interaction chromatography, and size exclusion chromatography. Furthermore, the Center provides assistance with biophysical characterization of proteins and their interactions with different types of molecules.
Ravi Kalathur, PhD