Orientation of antigen binding sites in dimeric and trimeric single chain Fv antibody fragments

FEBS Letters

Tulloch, Peter A.¹  Iliades, Peter²  Hudson, Peter J.²  Kortt, Alexander A.²  Lawrence, Lynne J.¹ 

[1] Biomolecular Research Institute, 343 Royal Parade, Parkville, Vic. 3052, Australia;CSIRO Molecular Science and CRC for Diagnostic Technologies, 343 Royal Parade, Parkville, Vic. 3052, Australia

关键词: Antibody;    Dimer;    Trimer;    Diabody;    Triabody;    Single chain Fv;    Antigen complex;    FPLC;    fast protein liquid chromatography;    IPTG;    isopropyl-β-thiogalactoside;    Fab;    antibody fragment produced by proteolysis;    Fv;    complex of VH and VL domains;    NA;    influenza neuraminidase;    M r;    molecular mass;    PCR;    polymerase chain reaction;    scFv;    single chain Fv molecule;    SDS-PAGE;    electrophoresis in a 15% polyacrylamide gel containing 1% SDS;    VH;    variable region from antibody heavy chain;    VL;    variable region from antibody light chain;    YT;    yeast tryptone medium;   

DOI  :  10.1016/S0014-5793(98)00292-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Electron microscopy of dimeric and trimeric single chain antibody Fv fragments (scFvs) complexed with anti-idiotype Fab fragments was used to reveal the orientation of antigen binding sites. This is the first structural analysis that discloses the multivalent binding orientation of scFv trimers (triabodies). Three different scFv molecules were used for the imaging analysis; NC10 scFv-5 and scFv-0, with five- and zero-residue linkers respectively between the V_H and V_L domains, were complexed with 3-2G12 anti-idiotype Fab fragments and 11-1G10 scFv-0 was complexed with NC41 anti-idiotype Fab fragments. The scFv-5 molecules formed bivalent dimers (diabodies) and the zero-linker scFv-0 molecules formed trivalent trimers (triabodies). The images of the NC10 diabody-Fab complex appear as boomerangs, not as a linear molecule, with a variable angle between the two Fab arms and the triabody-Fab complexes appear as tripods.

【 授权许可】

Unknown