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KMID : 0620920130450090004
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Experimental & Molecular Medicine
2013 Volume.45 No. 9 p.4 ~ p.0
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Application of bispecific antibody against antigen and hapten for immunodetection and immunopurification
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Kim Hyo-Ri
Park Sun-Young
Lee Hwa-Kyoung
Chung Jun-Ho
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Abstract
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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 × 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.
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KEYWORD
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bispecific antibody, hapten, immunoblot, immunoprecipitation
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