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Upon heating, the ricin binder still retained its specificity showing insignificantly binding to irrelevant targets, CT and SEB (Fig. 4B).
Both BoNT/A binders showed considerable cross-reactivity to other BoNT subtypes; this binding was significantly greater than binding to irrelevant targets (preliminary data not shown).
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Nonetheless, for the BoNT binders selected, both had stronger binding to the three tested BoNT complex toxoid subtypes, E, B, and A than to irrelevant targets (Fig. 3B).
However, we also observed relatively high binding to irrelevant control proteins (hen egg white ovalbumin and human apolipoprotein A1), so these data should be interpreted with some caution.
In all analyses, sdAb binding to irrelevant antigens was also monitored to ensure signal was not due to non-specific binding of unfolded sticky protein.
P1SD3-3 bound to SEB specifically relative to irrelevant targets, while P2SC8 exhibited more non-specific sticky interactions with irrelevant targets, CT and ricin (Figs 3D and 3E).
It seems to work by binding to multiple targets, she said, which may slow down the development of resistance.
Promiscuous binding to several protease targets demonstrates the emerging importance of quantitative studies on binding specificity.
Background binding to an irrelevant human IgG, MonoD, was subtracted.
Phage rescued from these clones were tested for binding to the specific RNA target and an irrelevant RNA target.
Controls are conducted in parallel (e.g., secondary antibody alone to check for non-specific binding to the replica, irrelevant primary antibody etc).
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