Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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B3(Fab)-streptavidin Tetramer Has Higher Binding Avidity than B3(scFv)-streptavidin Tetramer

  • Won, Jae-Seon (Collegeof Life Sciences and Graduate School of Biotechnology, Korea University) ;
  • Kang, Hye-Won (Collegeof Life Sciences and Graduate School of Biotechnology, Korea University) ;
  • Nam, Pil-Won (Collegeof Life Sciences and Graduate School of Biotechnology, Korea University) ;
  • Choe, Mu-Hyeon (Collegeof Life Sciences and Graduate School of Biotechnology, Korea University)
  • Published : 2009.05.20
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Abstract

Multivalent and multi-specific antibodies can provide valuable tools for bio-medical research, diagnosis and therapy. In antigen-antibody interactions, the avidity of antibodies depends on the affinity and the number of binding sites.$^1$ As artificial multivalent antibody agents, single chain Fv-streptavidin fusion tetramer proteins $(scFv-SA)_4$ have been previously tested.$^{1,\;2}$ Although, the Fab domain is known to be more stable than scFv in animal models,$^{3,\;4}$ it has never been used to make a multivalent agent with a streptavidin fusion. In this study, we prepared tetra-valent $(Fab-cSA)_4$ by fusing Fab with core streptavidin (cSA). This molecule was made using inclusion body production, refolding and chromatography purification. Affinities of the Fab-cSA tetramer and a scFv-cSA tetramer to a cell surface antigen were compared by ELISA using biotin-HRP. The Fab-cSA tetramer showed higher binding avidity than the scFv-cSA tetramer. The higher binding avidity of the Fab-cSA tetramer demonstrates its potential as a therapeutic agent for target-specific antibody therapy.

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References

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