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Gene Synthesis> | MATERIALS AND METHODS Construction of bispecific tandem scFv-Fc fusion vector A gene encoding the leader sequence of the human Ig k-chain, anti- complement C5 scFv (H Kim, HK Lee, WJ Yang, J Chung, unpublished data), a linker consisting of (Gly-Gly-Gly-Ser)3, anti- cotinine scFv,4 the hinge region of human IgG1 and the CH2–CH3 domains of rabbit IgG was chemically synthesized (Genscript, Piscataway, NJ, USA) with KpnI and BamHI restriction sites at the 50 and 30 ends, respectively. | Get A Quote |
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 c... More