MALDI-TOF-Based Affinity Selection Mass Spectrometry for Automated Screening of Protein–Ligand Interactions at High Throughput

High-Throughput Affinity Selection Mass Spectrometry Using SAMDI-MS to Identify Small-Molecule Binders of the Human Rhinovirus 3C Protease

“…The >100,000 compounds screened here feature an average purity >95%, and >95% of the compounds can be observed by MALDI MS in the positive reflector mode (data not shown), in line with previous results using MALDI. 7 The area under the curve (AUC) of each peak that corresponds to the mass ID (mass tolerance m / z 0.5) in the provided input sheet is measured from the raw spectra for each well on the SAMDI array and reported along with the AUC of the tri(ethylene glycol) (EG3) terminated monolayer molecule, which serves as an internal comparator having an m / z 335.2. All masses generate a nonzero AUC value given that the baseline in the MS spectra is never zero.…”

“…Currently, a common option is to infer hit identification from MS detection following several steps, including mixing the target with up to 2500 compounds, 6 isolating the complexes under native conditions, dissociation of the bound compounds through denatured conditions, and MS analysis. A recent report described an alternative workflow where complex isolation is performed in a 384-well filter plate assay format, followed by compound detection using high-throughput matrix-assisted laser desorption ionization (MALDI) MS. 7 Our team previously highlighted how the combination of self-assembled monolayers of alkanethiolates on gold and MALDI time-of-flight (TOF) MS, a technique termed self-assembled monolayer desorption ionization (SAMDI)–ASMS, can be used to identify small-molecule noncovalent binders. 8 The original proof-of-concept study showcased how SAMDI-ASMS enables affinity capture of a target, followed by subsequent ionization to infer the identification of the noncovalent compound by its mass.…”

An ultrafast and flexible liquid chromatography/tandem mass spectrometry system paves the way for machine learning driven in vivo sample processing in early drug discovery

“…RapidFire (RF) systems provide a medium‐throughput capacity with cycle times of 7–20 s, including a quick solid‐phase enrichment/cleanup step 1–6 or in BLAZE‐mode 2.5 s via direct injection 7 . Matrix‐assisted laser desorption/ionization time‐of‐flight (MALDI‐TOF) MS with a sample throughput of up to 0.4 s/sample 8–11 and acoustic droplet ejection open port interface (ADE‐OPI) MS with up to 6 samples/s are pushing the speed limits even further 12–15 …”

“…RapidFire (RF) systems provide a medium-throughput capacity with cycle times of 7-20 s, including a quick solid-phase enrichment/ cleanup step [1][2][3][4][5][6] or in BLAZE-mode 2.5 s via direct injection. 7 Matrixassisted laser desorption/ionization time-of-flight (MALDI-TOF) MS with a sample throughput of up to 0.4 s/sample [8][9][10][11] and acoustic droplet ejection open port interface (ADE-OPI) MS with up to 6 samples/s are pushing the speed limits even further. [12][13][14][15] Over the years, the RF-based initial Rapid and Integrated Analysis System (RIAS) was proven to be suitable for measuring samples from in vitro studies.…”

An Ultrafast and Flexible LC-MS/MS System Paves the Way for Machine Learning Driven Sample Processing and Data Evaluation in Early Drug Discovery

“…RapidFire (RF) systems provide a medium throughput capacity with cycle times of 7-20 s, including a quick solid-phase enrichment/cleanup step [1][2][3][4][5][6] or in BLAZE-mode 2.5 s via direct injection. 7 Matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) MS with a sample throughput of up to 0.4 s/sample [8][9][10][11] and acoustic droplet ejection open port interface (ADE-OPI) MS with up to 6 samples/s are pushing the speed limits even further. [12][13][14][15] Over the years, for the measurement of samples from in vitro studies, the RF-based initial RIAS system proved the suitability for this type of samples 5,6 .…”