scholarly journals High-throughput identification of proteins with AMPylation using self-assembled human protein (NAPPA) microarrays

2015 ◽  
Vol 10 (5) ◽  
pp. 756-767 ◽  
Author(s):  
Xiaobo Yu ◽  
Joshua LaBaer
Keyword(s):  
High Throughput ◽  
Human Protein ◽  
Self Assembled

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

2021 ◽  
pp. 247255522110232
Author(s):  
Michael D. Scholle ◽  
Doug McLaughlin ◽  
Zachary A. Gurard-Levin
Keyword(s):  
Mass Spectrometry ◽  
Drug Discovery ◽  
High Throughput ◽  
High Throughput Screening ◽  
Human Rhinovirus ◽  
Binding Potential ◽  
Affinity Selection ◽  
Response Curves ◽  
Desorption Ionization ◽  
Self Assembled

Affinity selection mass spectrometry (ASMS) has emerged as a powerful high-throughput screening tool used in drug discovery to identify novel ligands against therapeutic targets. This report describes the first high-throughput screen using a novel self-assembled monolayer desorption ionization (SAMDI)–ASMS methodology to reveal ligands for the human rhinovirus 3C (HRV3C) protease. The approach combines self-assembled monolayers of alkanethiolates on gold with matrix-assisted laser desorption ionization time-of-flight (MALDI TOF) mass spectrometry (MS), a technique termed SAMDI-ASMS. The primary screen of more than 100,000 compounds in pools of 8 compounds per well was completed in less than 8 h, and informs on the binding potential and selectivity of each compound. Initial hits were confirmed in follow-up SAMDI-ASMS experiments in single-concentration and dose–response curves. The ligands identified by SAMDI-ASMS were further validated using differential scanning fluorimetry (DSF) and in functional protease assays against HRV3C and the related SARS-CoV-2 3CLpro enzyme. SAMDI-ASMS offers key benefits for drug discovery over traditional ASMS approaches, including the high-throughput workflow and readout, minimizing compound misbehavior by using smaller compound pools, and up to a 50-fold reduction in reagent consumption. The flexibility of this novel technology opens avenues for high-throughput ASMS assays of any target, thereby accelerating drug discovery for diverse diseases.

scholarly journals Development of a Novel Label-Free and High-Throughput Arginase-1 Assay Using Self-Assembled Monolayer Desorption Ionization Mass Spectrometry

2021 ◽  
pp. 247255522110006
Author(s):  
Michael D. Scholle ◽  
Zachary A. Gurard-Levin
Keyword(s):  
Mass Spectrometry ◽  
Drug Discovery ◽  
High Throughput ◽  
Ionization Mass Spectrometry ◽  
Ionization Mass ◽  
Label Free ◽  
Desorption Ionization ◽  
Arginase 1 ◽  
Self Assembled ◽  
High Throughput Assay

Arginase-1, an enzyme that catalyzes the reaction of L-arginine to L-ornithine, is implicated in the tumor immune response and represents an interesting therapeutic target in immuno-oncology. Initiating arginase drug discovery efforts remains a challenge due to a lack of suitable high-throughput assay methodologies. This report describes the combination of self-assembled monolayers and matrix-assisted laser desorption ionization mass spectrometry to enable the first label-free and high-throughput assay for arginase activity. The assay was optimized for kinetically balanced conditions and miniaturized, while achieving a robust assay (Z-factor > 0.8) and a significant assay window [signal-to-background ratio > 20] relative to fluorescent approaches. To validate the assay, the inhibition of the reference compound nor-NOHA (Nω-hydroxy-nor-L-arginine) was evaluated, and the IC50 measured to be in line with reported results (IC50 = 180 nM). The assay was then used to complete a screen of 175,000 compounds, demonstrating the high-throughput capacity of the approach. The label-free format also eliminates opportunities for false-positive results due to interference from library compounds and optical readouts. The assay methodology described here enables new opportunities for drug discovery for arginase and, due to the assay flexibility, can be more broadly applicable for measuring other amino acid–metabolizing enzymes.

scholarly journals High throughput trapping and arrangement of biological cells using self-assembled optical tweezer

2018 ◽  
Vol 26 (26) ◽  
pp. 34665 ◽  
Author(s):  
Zongbao Li ◽  
Jianxin Yang ◽  
Shaojing Liu ◽  
Xiaofang Jiang ◽  
Haiyan Wang ◽  
...  
Keyword(s):  
High Throughput ◽  
Optical Tweezer ◽  
Biological Cells ◽  
Self Assembled

scholarly journals High-throughput photocapture approach for reaction discovery

2020 ◽  
Vol 117 (24) ◽  
pp. 13261-13266 ◽  
Author(s):  
Alison A. Bayly ◽  
Benjamin R. McDonald ◽  
Milan Mrksich ◽  
Karl A. Scheidt
Keyword(s):  
High Throughput ◽  
Rapid Assessment ◽  
Ionization Mass ◽  
Reductive Coupling ◽  
Self Assembled Monolayer ◽  
High Throughput Analysis ◽  
Reaction Discovery ◽  
Assembled Monolayers ◽  
Self Assembled ◽  
Bioactive Agents

Modern organic reaction discovery and development relies on the rapid assessment of large arrays of hypothesis-driven experiments. The time-intensive nature of reaction analysis presents the greatest practical barrier for the execution of this iterative process that underpins the development of new bioactive agents. Toward addressing this critical bottleneck, we report herein a high-throughput analysis (HTA) method of reaction mixtures by photocapture on a 384-spot diazirine-terminated self-assembled monolayer, and self-assembled monolayers for matrix-assisted laser desorption/ionization mass spectrometry (SAMDI-MS) analysis. This analytical platform has been applied to the identification of a single-electron-promoted reductive coupling of acyl azolium species.

Ligand identification of the adenosine A2Areceptor in self-assembled nanodiscs by affinity mass spectrometry

2017 ◽  
Vol 9 (40) ◽  
pp. 5851-5858 ◽  
Author(s):  
Jun Ma ◽  
Yan Lu ◽  
Dong Wu ◽  
Yao Peng ◽  
Wendy Loa-Kum-Cheung ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
High Throughput Screening ◽  
Ligand Identification ◽  
Self Assembled

The combination of nanodisc and affinity LC/MS techniques potentiates the high-throughput screening against GPCRs.

Sign in / Sign up

Export Citation Format

Share Document