MS Binding Assays

Benocyclidine (BTCP) as Non‐labelled Reporter Ligand for MS Binding Assays for the PCP Ion Channel Binding Site of the Desensitized Torpedo Nicotinic Acetylcholine Receptor (nAChR)

“…As for the quantification of analytes in biological matrices also the quantification of reporter ligands in MS Binding Assays [28,29] by LC-ESI-MS/MS may be accomplished employing an external or internal standard. However, the quantification based on an internal standard can distinctly improve the robustness of a LC-ESI-MS quantification method, in particular when internal standard and analyte are virtually co-eluting.…”

A Library Screening Strategy Combining the Concepts of MS Binding Assays and Affinity Selection Mass Spectrometry

“…Despite these qualities, employment of “hot,” i.e., radioisotope labeled ligands is associated with disadvantages such as increased synthetic effort, hazards to human health, restrictions set by authorities, or expensive waste management. In order to overcome these limitations, several alternatives to avoid radioisotope labeled compounds, such as fluorescence, luminescence, surface plasmon resonance, or mass spectrometry (MS) based binding assays were established in the recent year (Geoghegan and Kelly, 2005; Zhu and Cuozzo, 2009; Stahelin, 2013; Höfner and Wanner, 2015; Stoddart et al, 2016). Although all of these alternatives provide specific strengths (and weaknesses) that should not be discussed in detail here, MS may be considered as particularly attractive detection principle as neither ligand nor target has to be modified or labeled for investigation of target-ligand interactions (Geoghegan and Kelly, 2005; Schermann et al, 2005; Höfner and Wanner, 2015).…”

“…In order to overcome these limitations, several alternatives to avoid radioisotope labeled compounds, such as fluorescence, luminescence, surface plasmon resonance, or mass spectrometry (MS) based binding assays were established in the recent year (Geoghegan and Kelly, 2005; Zhu and Cuozzo, 2009; Stahelin, 2013; Höfner and Wanner, 2015; Stoddart et al, 2016). Although all of these alternatives provide specific strengths (and weaknesses) that should not be discussed in detail here, MS may be considered as particularly attractive detection principle as neither ligand nor target has to be modified or labeled for investigation of target-ligand interactions (Geoghegan and Kelly, 2005; Schermann et al, 2005; Höfner and Wanner, 2015). Even in the field of MS based binding assays, a wealth of concepts has been reported covering direct measurement of target-ligand complexes (Hofstadler and Sannes-Lowery, 2006) as well as monitoring bound ligands after their liberation from the target (Annis et al, 2007a; Höfner and Wanner, 2015) or even recording of ligands remaining non-bound (Hofner and Wanner, 2003) in presence of the target.…”

“…Although all of these alternatives provide specific strengths (and weaknesses) that should not be discussed in detail here, MS may be considered as particularly attractive detection principle as neither ligand nor target has to be modified or labeled for investigation of target-ligand interactions (Geoghegan and Kelly, 2005; Schermann et al, 2005; Höfner and Wanner, 2015). Even in the field of MS based binding assays, a wealth of concepts has been reported covering direct measurement of target-ligand complexes (Hofstadler and Sannes-Lowery, 2006) as well as monitoring bound ligands after their liberation from the target (Annis et al, 2007a; Höfner and Wanner, 2015) or even recording of ligands remaining non-bound (Hofner and Wanner, 2003) in presence of the target. In the context of the present study, we want to focus on two strategies of MS based binding assays, namely MS Binding Assays (Höfner and Wanner, 2015; Massink et al, 2015) and affinity selection mass spectrometry (ASMS) (Van Breemen et al, 1997; Zehender et al, 2004; Annis et al, 2007a; Jonker et al, 2011).…”

“…Even in the field of MS based binding assays, a wealth of concepts has been reported covering direct measurement of target-ligand complexes (Hofstadler and Sannes-Lowery, 2006) as well as monitoring bound ligands after their liberation from the target (Annis et al, 2007a; Höfner and Wanner, 2015) or even recording of ligands remaining non-bound (Hofner and Wanner, 2003) in presence of the target. In the context of the present study, we want to focus on two strategies of MS based binding assays, namely MS Binding Assays (Höfner and Wanner, 2015; Massink et al, 2015) and affinity selection mass spectrometry (ASMS) (Van Breemen et al, 1997; Zehender et al, 2004; Annis et al, 2007a; Jonker et al, 2011). MS Binding Assays are closely related to radioligand binding assays and share a lot of common features such as setup of the binding experiment as well as concentrations of ligands and targets.…”