This detection limitation prompted the development of a FAC-MS method, which significantly expands the scope of the method to complex mixtures of compounds. An MS approach removes the requirement for labelling compounds to enhance a fluorescent signal and minimizes the need to pre-purify the samples to be analyzed. Monitoring numerous compounds via their respective m/z values enables the determination of individual breakthrough curves from mixtures and offers powerful insights into multi-ligand behavior. At the simplest level, this combination of technologies (FAC, MS) provides the opportunity to rank-order binding strength in a single experiment, immediately placing the discovery of new ligands in a relational context. However, the opportunity to monitor multiple breakthrough curves without ligand labelling presents additional advantages. To illustrate, we will consider two different classes of methods: direct and indirect.
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