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. 2016 Sep 8;6:32747. doi: 10.1038/srep32747

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Copyright © 2016, The Author(s)

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(A) Example of fluid-phase affinity measurement of TNF-anti-TNF. DyLight-488 labeled adalimumab Fab’ is incubated with different concentrations of TNF and analyzed by HP-SEC, monitoring the elution of bound and free adalimumab Fab’-488 by in-line fluorescence detection. (B) Free Fab-488 is plotted vs the concentration of TNF (molar concentration of binding sites) and a 1:1 Langmuir binding model is fitted, thus assuming independent association of anti-TNF Fab’s to each binding site on TNF. (C) TNF-488 was incubated with different concentrations of etanercept and analyzed analogously to B, using the molar concentration of etanercept in the calculations. Representative of n = 3. F = fluorescence.