Production and characterization of anti-human interferon {gamma} receptor antibody fragments that inhibit cytokine binding to the receptor |
| |
Authors: | Bridges Angela; Stuart Fiona; Spth Julia; Lang Stefan; Henke Christoph; Birch Ashley; Robinson John A |
| |
Affiliation: | Institute of Organic Chemistry, University of Zurich Winterthurerstrasse 190, 8057 Zurich, Switzerland |
| |
Abstract: | Three single-chain antibody fragments that recognize the extracellularhuman interferon receptor -chain (IFN R), and inhibit the bindingof human IFN , have been produced in Escherichia coli. Thesefragments are derived from murine anti-receptor monoclonal antibodies,and comprise the variable heavy (VH) domain linked to the variablelight (VL) chain through a 15 amino acid linker (GGGGS)3].Using surface plasmon resonance technology (BIAcore), the solubleproteins were shown to retain a high affinity for recombinantIFN R, and by radioimmunoassay to possess high inhibitory activitytowards IFN -binding to human Raji cells. The antibody fragmentsmost likely recognize epitopes that overlap the cytokine bindingsite on the receptor surface. Attempts to dissect further theantibodies to isolated VH- and VL-chains and to synthetic linearand cyclic peptides derived from the individual complementaritydetermining regions failed to afford fragments with significantIFN R binding affinity. Nevertheless, these native-like variableregion fragments and petidomimetics derived from them are ofinterest in the design of novel IFN R antagonists. |
| |
Keywords: | antagonist/ epitope/ Fv fragment/ ligand design/ mimetic/ monoclonal antibody |
本文献已被 Oxford 等数据库收录! |
|