Nathalie Plundrich, Mary Ann Lila, Edward Foegeding, Scot Laster (2017). Protein-bound polyphenols create “ghost” band artifacts during chemiluminescence-based antigen detection. F1000 Research 6(254).
Plants for Human Health Institute , North Carolina Research Campus, North Carolina State University
Department of Food, Bioprocessing and Nutrition Sciences, North Carolina State University, North Carolina, USA
Department of Biological Sciences, North Carolina State University, North Carolina, USA
Antigen detection during Western blotting commonly utilizes a horseradish
peroxidase-coupled secondary antibody and enhanced chemiluminescent
substrate. We utilized this technique to examine the impact of green
tea-derived polyphenols on the binding of egg white protein-specific IgE
antibodies from allergic human plasma to their cognate antigens. Our
experiments unexpectedly showed that green tea-derived polyphenols, when
stably complexed with egg white proteins, caused hyperactivation of
horseradish peroxidase resulting in the appearance of white “ghost” bands.
This study suggests that caution should be taken when evaluating
polyphenol-bound proteins by enhanced chemiluminescence Western blotting
using horseradish peroxidase and demonstrates that protein-bound
polyphenols can be a source of “ghost” band artifacts on Western blots.