Conversion of scFv peptide-binding specificity for crystal chaperone development.

ABSTRACT: In spite of advances in protein expression and purification over the last decade, many proteins remain recalcitrant to structure determination by X-ray crystallography. One emerging tactic to obtain high-quality protein crystals for structure determination, particularly in the case of membrane proteins, involves co-crystallization with a protein-specific antibody fragment. Here, we report the development of new recombinant single-chain antibody fragments (scFv) capable of binding a specific epitope that can be introduced into internal loops of client proteins. The previously crystallized hexa-histidine-specific 3D5 scFv antibody was modified in the complementary determining region and by random mutagenesis, in conjunction with phage display, to yield scFvs with new biochemical characterist

SUBMITTER: Pai JC 

PROVIDER: S-EPMC3077810 | biostudies-literature | 2011 May

REPOSITORIES: biostudies-literature