Generation of Synthetic Antibody Fragments to Detergent Solubilized Membrane Proteins

Uysal S, Kossiakoff AA

Springer Protocols Handbook, p1-13


Structural determination of membrane proteins is extremely challenging due to the physical characteristics of membrane proteins themselves and the lack of adequate tools and technologies to perform the studies. Recent developments in micro-focus X-ray beams, novel detergents, protein thermo-stabilization, and protein engineering have been essential in expanding the pool of membrane proteins deposited in PDB. Despite these advances, crystallization of membrane proteins still remains the main bottleneck in obtaining high quality structures. Recently, the use of antibody and non-antibody scaffold binding partners as crystallization “chaperones” has emerged as a powerful method to obtaining well-diffracting crystals of membrane proteins. In this chapter, a protocol is provided to generate synthetic antibody fragments for use as crystallization chaperones for membrane proteins.