Production and characterization of anti-human interferon gamma receptor antibody fragments that inhibit cytokine binding to the receptor.
Three single-chain antibody fragments that recognize the extracellular human interferon gamma receptor alpha-chain (IFN gamma R), and inhibit the binding of human IFN gamma, have been produced in Escherichia coli. These fragments are derived from murine anti-receptor monoclonal antibodies, and comprise the variable heavy (VH) domain linked to the variable light (VL) chain through a 15 amino acid linker [(GGGGS)3]. Using surface plasmon resonance technology (BIAcore), the soluble proteins were shown to retain a high affinity for recombinant IFN gamma R, and by radioimmunoassay to possess a high inhibitory activity towards IFN gamma-binding to human Raji cells. The antibody fragments most likely recognize epitopes that overlap the cytokine binding site on the receptor surface. Attempts to dissect further the antibodies to isolated VH- and VL-chains and to synthetic linear and cyclic peptides derived from the individual complementarity determining regions failed to afford fragments with significant IFN gamma R binding affinity. Nevertheless, these native-like variable region fragments and petidomimetics derived from them are of interest in the design of novel IFN gamma R antagonists.
Bridges
A
A
Institute of Organic Chemistry, University of Zurich, Switzerland.
Stuart
F
F
Späth
J
J
Lang
S
S
Henke
C
C
Birch
A
A
Robinson
J A
JA
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000494:pharmacology*; D015703:Antigens, CD / Q000276:immunology*; D001483:Base Sequence; D015374:Biosensing Techniques; D000939:Epitopes; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000494:pharmacology*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000494:pharmacology; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000494:pharmacology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000494:pharmacology; D007371:Interferon-gamma / Q000378:metabolism*; D008969:Molecular Sequence Data; D011863:Radioimmunoassay; D017471:Receptors, Interferon / Q000276:immunology*; D011994:Recombinant Proteins / Q000494:pharmacology; D000096743:Interferon gamma Receptor
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000939:Epitopes; D007128:Immunoglobulin Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D007371:Interferon-gamma; D017471:Receptors, Interferon; D011994:Recombinant Proteins; D000096743:Interferon gamma Receptor; C055966:immunoglobulin Fv