DESIGN: Computerized optimization of experimental design for estimating Kd and Bmax in ligand binding experiments: I. Homologous and heterologous binding to one or two classes of sites
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Evaluation of the Flow-Dialysis Technique for Analysis of Protein-Ligand Interactions: An Experimental and a Monte Carlo Study
2004, Biophysical JournalCitation Excerpt :This approach enabled us to accurately determine both the KD and E0 with one sample in a single experiment within ∼30 min. Several reports in the literature have indicated that improper analysis of the obtained data (e.g., ignoring background signal, overlooking nonspecific binding, a nonlinear relationship between a measured signal and theconcentration of bound or free ligand, or the presence of partially inactivated components) and the use of inappropriate mathematical procedures to evaluate the data can cause severe bias in the obtained ligand binding parameters (Fuchs and Gessner, 2001; Larsson, 1997; Rovati et al., 1988; Schumacher and Von Tscharner, 1994). Thus, although flow dialysis is experimentally a simple and fast technique, accurate determination of the protein-ligand parameters necessitates a thorough evaluation of the experimental procedure.
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