Development of a High-Throughput Screening Assay to Identify Inhibitors of the Major M17-Leucyl Aminopeptidase from Trypanosoma cruzi Using RapidFire Mass Spectrometry

Expression in Escherichia coli, purification and kinetic characterization of LAPLm, a Leishmania major M17-aminopeptidase

“…LAPLm appears to be involved in nutritional supply, since the parasite lacks the biosynthetic pathways for branched side chain amino acids, including leucine [23]. Consistent with this crucial role, arphamenine A, a Trypanosoma cruzi acidic M17 LAP inhibitor [24], inhibits in vitro growth of related T. brucei brucei [12], suggesting functional conservation across kinetoplastids. Down-regulation of TbLAP1, a T. brucei M17 LAP involved in mitochondrial kinetoplast DNA segregation during cell division, generates a cytokinesis delay [16].…”

Automated High-Throughput System Combining Small-Scale Synthesis with Bioassays and Reaction Screening

“…Solid-phase extraction (SPE)-based platforms (e.g., Agilent RapidFire), which provide online enrichment and sample cleanup, reducing ionization suppression and matrix effects but increasing analysis time (~8 s/sample). 13,41,42,53–55…”

Recent advances in analytical techniques for high throughput experimentation

“…In recent years, many industries and scientific disciplines have embraced the concept of high throughput experimentation (HTE) to support the ever-increasing need for faster data generation and acceleration of product development cycles. In pharmaceutical industry, HTE has been adopted in diverse areas, such as the discovery of biomarkers and of new chemical entities in drug discovery, [1][2][3] as a ABBREVIATIONS: ADE, acoustic droplet ejection; AEMS, acoustic ejection mass spectrometry; AIMS, ambient ionization mass spectrometry; AMI, acoustic mist ionization; AP, atmospheric pressure; APCI, atmospheric-pressure chemical ionization; APPI, atmospheric-pressure photoionization; BPR, back pressure regulator; CD, circular dichroism; DART, direct analysis in real-time; DESI, desorption electrospray ionization; FPP, fully porous particles; HTA, high throughput analysis; HTE, high throughput experimentation; HTS, high throughput screening; IMS, ion mobility spectrometry; MISER, multiple injections in a single experimental run; OPI, open port interface; OT, open tubular; SAMDI, self-assembled monolayers coupled with desorption/ionization; SFC, supercritical fluid chromatography; SIMS, secondary ion mass spectrometry; SPME, solid-phase microextraction; SPP, superficially porous particles; UHPLC, ultrahigh-performance liquid chromatography/ultrahigh-pressure liquid chromatography; UV, ultraviolet This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made. © 2021 The Authors.…”

“…In recent years, many industries and scientific disciplines have embraced the concept of high throughput experimentation (HTE) to support the ever‐increasing need for faster data generation and acceleration of product development cycles. In pharmaceutical industry, HTE has been adopted in diverse areas, such as the discovery of biomarkers and of new chemical entities in drug discovery, 1 , 2 , 3 as a tool to accelerate the characterization of new pharmaceutical compounds as well as small‐molecule chemical process development, 4 , 5 , 6 , 7 in biotherapeutics analysis, 8 forced degradation studies of therapeutic peptides, 9 and analytical method qualification. 10 …”