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Experimental & Molecular Medicine ; : e43-2013.
Artículo en Inglés | WPRIM | ID: wpr-71812

RESUMEN

We present a bispecific antibody that recognizes an antigen and a hapten and can be applied to various biological assays, including immunoblotting and immunoprecipitation. In immunoblot analysis of serum, an anti-C5 x anti-cotinine bispecific tandem single-chain variable fragment (scFv)-Fc fusion protein and cotinine-conjugated horseradish peroxidase (HRP) generated a clean signal without the high background that was observed in a parallel experiment using HRP-conjugated goat anti-rabbit immunoglobulin G (Fc-specific) antibody. In immunoprecipitation analysis of serum, use of the bispecific tandem scFv-Fc fusion protein and cotinine-crosslinked magnetic beads significantly reduced the amount of protein contaminants compared with a parallel experiment done with protein A agarose beads. In subsequent immunoblot analysis, use of cotinine-HRP as the secondary probe instead of HRP-conjugated goat anti-rabbit IgG (Fc-specific) antibody successfully eliminated the band corresponding to the bispecific tandem scFv-Fc fusion protein.


Asunto(s)
Animales , Humanos , Conejos , Anticuerpos Biespecíficos/inmunología , Células HEK293 , Haptenos/inmunología , Immunoblotting/métodos , Inmunoprecipitación/métodos , Anticuerpos de Cadena Única/inmunología
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