Label-free high-throughput assays to screen and characterize novel lactate dehydrogenase inhibitors

Measurement of glycolysis reactants by high-throughput solid phase extraction with tandem mass spectrometry: Characterization of pyrophosphate-dependent phosphofructokinase as a case study

“…The MS-based analysis of multienzyme glycolytic systems has been described previously [28], further suggesting that high-throughput SPE-MS/MS could be instrumental in simultaneously studying multiple enzymes within a complex network. Because SPE-MS/MS can accommodate a variety of packing materials, can be run in reverse of normal phase, and can be connected to any mass spectrometer, this detection strategy facilitates the measurement of a wide breadth of analytes including small molecules [29][30][31][32][33][34], peptides [35][36][37], and whole proteins [38]. This work, therefore, also contributes to a growing list of applications for high-throughput SPE-MS/MS analysis.…”

Plasmodial enzymes in metabolic pathways as therapeutic targets and contemporary strategies to discover new antimalarial drugs: a review

“…During a typical HTS assay, a collection of chemical composites is carefully chosen and tested against a biological target to evaluate the strength of the related inhibition or activation signal (Malo et al, 2006). In order to successfully identify LDH inhibitors from library of a small molecule compound, two label-free high-throughput assays were planned using a kinetic high-throughput screen (VanderPorten et al, 2013). In addition, examination on antimalarial elements from Berberis thunbergii and Eugenia rigida by automated HTS, using UPLC-MS-ELSD-PDA yielded two triterpenoids, namely α-betulinic acid and β-betulinic acid, that were acknowledged as antimalarial active elements from HTS hits of E. rigida (Zhang et al, 2014) High throughput screening is not only beneficial in the search for antimalarial drugs but is also indispensable in the discovery of small molecules that can interfere malaria transmission.…”

Biofuel metabolic engineering with biosensors

“…However, these methods require time- and labor-intensive metabolite extractions that result in destructive, bulk measurements that are generally low throughput (10 1 –10 3 per day). Two emerging MS-based platforms that may aid in overcoming these limitations are the RapidFire high-throughput MS system from Agilent Technology, Inc. [7] and surface-based MS techniques, such as Nanostructure-Initiator MS (NIMS) [8]. …”

“…Samples in microtiter plates are purified by solid-phase extraction and directly injected into an MS instrument. The instrument can processes a single sample in less than 15 s, which is over 100x faster than traditional liquid-chromatography-MS measurements [7]. NIMS is a surface-assisted laser desorption/ionization technique that requires little sample preparation and uses a liquid “initiator,” instead of a co-crystallization matrix, to produce spectra with high sensitivity and lower noise in the metabolite mass region.…”