MS Binding Assays—with MALDI toward High Throughput

Höfner, Georg; Merkel, Dietrich; Wanner, Klaus T.

doi:10.1002/cmdc.200900201

Cited by 16 publications

(11 citation statements)

References 29 publications

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“…This technology has been used to determine the binding of small molecules to the murine GABA (gamma-aminobutyric acid) receptor mGAT 1 [87], to determine the inhibition of protein kinase A (PKA) [88], and, in a multiplexed system, to investigate the enzyme activity of a kinase and an esterase in a single analysis [89]. …”

Section: Off-line and At-line Bioaffinity Analysis With Parallel Ms Dmentioning

confidence: 99%

Advances in mass spectrometry-based post-column bioaffinity profiling of mixtures

Kool¹,

Giera²,

Irth³

et al. 2010

Anal Bioanal Chem

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In the screening of complex mixtures, for example combinatorial libraries, natural extracts, and metabolic incubations, different approaches are used for integrated bioaffinity screening. Four major strategies can be used for screening of bioactive mixtures for protein targets—pre-column and post-column off-line, at-line, and on-line strategies. The focus of this review is on recent developments in post-column on-line screening, and the role of mass spectrometry (MS) in these systems. On-line screening systems integrate separation sciences, mass spectrometry, and biochemical methodology, enabling screening for active compounds in complex mixtures. There are three main variants of on-line MS based bioassays: the mass spectrometer is used for ligand identification only; the mass spectrometer is used for both ligand identification and bioassay readout; or MS detection is conducted in parallel with at-line microfractionation with off-line bioaffinity analysis. On the basis of the different fields of application of on-line screening, the principles are explained and their usefulness in the different fields of drug research is critically evaluated. Furthermore, off-line screening is discussed briefly with the on-line and at-line approaches.Schematic view of an on-line bioaffinity analysis or HRS setup with MS based bioassay detection

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Section: Off-line and At-line Bioaffinity Analysis With Parallel Ms Dmentioning

confidence: 99%

Advances in mass spectrometry-based post-column bioaffinity profiling of mixtures

Kool¹,

Giera²,

Irth³

et al. 2010

Anal Bioanal Chem

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“…A followup technology used MALDI as an ionization source. This adaptation allowed a large throughput increase owing to rapid analysis of the eventually spotted 96-sample MALDI plates [72]. Slightly different approaches use a rapid filtration step (in 96-filtration well plates) instead of SEC to obtain the same assay principle.…”

Section: Affinity Selection—binding To Targets In Solutionmentioning

confidence: 99%

Recent developments in protein–ligand affinity mass spectrometry

Jonker¹,

Kool²,

Irth³

et al. 2010

Anal Bioanal Chem

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This review provides an overview of direct and indirect technologies to screen protein–ligand interactions with mass spectrometry. These technologies have as a key feature the selection or affinity purification of ligands in mixtures prior to detection. Specific fields of interest for these technologies are metabolic profiling of bioactive metabolites, natural extract screening, and the screening of libraries for bioactives, such as parallel synthesis libraries and small combichem libraries. The review addresses the principles of each of the methods discussed, with a focus on developments in recent years, and the applicability of the methods to lead generation and development in drug discovery.FigureSchematic view of the principle of filtration based 96-well affinity selection MS binding assays

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“…The most straightforward conception solving this problem is to forgo chromatographic separation before MS quantification. Very recently we were able to demonstrate the feasibility of this analytical strategy employing a new MALDI-MS/MS system (FlashQuant) [9]. The measurement of a binding sample spotted onto a 96 well format MALDI plate took only 1.7 s, however, 14 s were required in total as the mean duration per sample.…”

Section: Introductionmentioning

confidence: 99%