Triabodies: single chain Fv fragments without a linker form trivalent trimers

FEBS Letters

Iliades, Peter¹  Hudson, Peter J¹  Kortt, Alexander A¹ 

[1] CSIRO, Division of Biomolecular Engineering, 343 Royal Parade, Parkville, Victoria 3052, Australia

关键词: Antibody;    Dimer;    Trimer;    Single chain Fv;    scFv;    Antigen complex;    BIAcore;    Pharmacia BIAcore 1000 biosensor apparatus;    FPLC;    fast protein liquid chromatography;    IPTG;    isopropyl-β thiogalactoside;    NA;    influenza neuraminidase;    M r;    molecular mass;    PCR;    polymerase chain reaction;    scFv;    single chain Fv molecule;    SDS-PAGE;    electrophoresis in a 15% polyacrylamide gel comprising 1% SDS;    RU;    resonance units;    VH;    variable region from antibody heavy chain;    VL;    variable region from antibody light chain;    YT;    yeast tryptone medium;   

DOI  :  10.1016/S0014-5793(97)00475-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A single chain Fv fragment (scFv) of the murine monoclonal antibody 11-1G10 was constructed by directly joining the C-terminal residue of the V_H domain to the N-terminal residue of V_L. 11-1G10 is an anti-idiotype and competes with the antigen, influenza virus neuraminidase (NA), for binding to the NC41 antibody. The scFv formed stable trimers with three active antigen combining sites for NC41 Fab fragments. We propose that trimeric scFvs may be the preferred conformation for directly linked V_H-V_L molecules, which contrasts the formation of scFv dimers (diabodies) when the V_H and V_L domains are joined by short flexible linkers of between 5–10 residues. BIAcore biosensor binding experiments showed that the trimeric scFv showed an expected increase in binding affinity, due to avidity, compared to the monomeric 15-residue linked scFv. The increase in avidity of scFv trimers offers advantages for imaging and immunotherapy.

【 授权许可】

Unknown