Metabolite fingerprinting by infrared matrix-assisted laser desorption electrospray ionization mass spectrometry
The adoption of mass spectrometry for high-throughput screening in drug discovery has become increasingly prevalent and has enabled label-free screening against diverse targets. Cellular assays for phenotypic screening, however, are primarily conducted by microscopy as there remain many challenges a...
Saved in:
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-06-01
|
| Series: | SLAS Technology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2472630325000305 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850047643338670080 |
|---|---|
| author | Alena N. Joignant Fan Pu Shaun M. McLoughlin James W. Sawicki Andrew J. Radosevich Renze Ma Jon D. Williams Sujatha M. Gopalakrishnan Nathaniel L. Elsen |
| author_facet | Alena N. Joignant Fan Pu Shaun M. McLoughlin James W. Sawicki Andrew J. Radosevich Renze Ma Jon D. Williams Sujatha M. Gopalakrishnan Nathaniel L. Elsen |
| author_sort | Alena N. Joignant |
| collection | DOAJ |
| description | The adoption of mass spectrometry for high-throughput screening in drug discovery has become increasingly prevalent and has enabled label-free screening against diverse targets. Cellular assays for phenotypic screening, however, are primarily conducted by microscopy as there remain many challenges associated with conducting phenotypic screens via ultra-high throughput mass spectrometry.Following a simple on-plate extraction, infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) was employed to directly sample the cell lysate at a speed of one sample per second with high mass resolution. A549 cells were treated with compounds identified as hits in literature, including a recently reported glutaminase cellular screen. Among the test compounds were confirmed glutaminase inhibitors, proposed nuisance compounds, and cell-active but enzyme-inactive compounds. Filtered data were further processed in R for dimensionality reduction and unsupervised clustering. The general nature of dimensionality reduction enables the immediate use of this method in applications other than glutaminase inhibition.Though we observed that all compounds affected the intracellular conversion of glutamine to glutamate, there were clear metabolic differences between the biochemically active compounds and the off-target false hits. Moreover, two nuisance compounds were observed to cluster separately from the confirmed glutaminase inhibitors in the observed metabolite fingerprints.This proof-of-concept work establishes a workflow that enables high-throughput mass spectrometry-based phenotypic screening. The methods proposed herein, at the throughput enabled by IR-MALDESI, could offer a new avenue for the discovery of novel drugs. |
| format | Article |
| id | doaj-art-3a59316846bf44fc9810cf958f12ffb0 |
| institution | DOAJ |
| issn | 2472-6303 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | SLAS Technology |
| spelling | doaj-art-3a59316846bf44fc9810cf958f12ffb02025-08-20T02:54:11ZengElsevierSLAS Technology2472-63032025-06-013210027210.1016/j.slast.2025.100272Metabolite fingerprinting by infrared matrix-assisted laser desorption electrospray ionization mass spectrometryAlena N. Joignant0Fan Pu1Shaun M. McLoughlin2James W. Sawicki3Andrew J. Radosevich4Renze Ma5Jon D. Williams6Sujatha M. Gopalakrishnan7Nathaniel L. Elsen8Discovery Research, AbbVie Inc., 1 North Waukegan Road, North Chicago, IL 60064, United StatesDiscovery Research, AbbVie Inc., 1 North Waukegan Road, North Chicago, IL 60064, United StatesDiscovery Research, AbbVie Inc., 1 North Waukegan Road, North Chicago, IL 60064, United StatesDiscovery Research, AbbVie Inc., 1 North Waukegan Road, North Chicago, IL 60064, United StatesDiscovery Research, AbbVie Inc., 1 North Waukegan Road, North Chicago, IL 60064, United StatesDiscovery Research, AbbVie Inc., 1 North Waukegan Road, North Chicago, IL 60064, United StatesDiscovery Research, AbbVie Inc., 1 North Waukegan Road, North Chicago, IL 60064, United StatesDiscovery Research, AbbVie Inc., 1 North Waukegan Road, North Chicago, IL 60064, United StatesCorresponding author at: Discovery Research, AbbVie Inc.; Discovery Research, AbbVie Inc., 1 North Waukegan Road, North Chicago, IL 60064, United StatesThe adoption of mass spectrometry for high-throughput screening in drug discovery has become increasingly prevalent and has enabled label-free screening against diverse targets. Cellular assays for phenotypic screening, however, are primarily conducted by microscopy as there remain many challenges associated with conducting phenotypic screens via ultra-high throughput mass spectrometry.Following a simple on-plate extraction, infrared matrix-assisted laser desorption electrospray ionization (IR-MALDESI) was employed to directly sample the cell lysate at a speed of one sample per second with high mass resolution. A549 cells were treated with compounds identified as hits in literature, including a recently reported glutaminase cellular screen. Among the test compounds were confirmed glutaminase inhibitors, proposed nuisance compounds, and cell-active but enzyme-inactive compounds. Filtered data were further processed in R for dimensionality reduction and unsupervised clustering. The general nature of dimensionality reduction enables the immediate use of this method in applications other than glutaminase inhibition.Though we observed that all compounds affected the intracellular conversion of glutamine to glutamate, there were clear metabolic differences between the biochemically active compounds and the off-target false hits. Moreover, two nuisance compounds were observed to cluster separately from the confirmed glutaminase inhibitors in the observed metabolite fingerprints.This proof-of-concept work establishes a workflow that enables high-throughput mass spectrometry-based phenotypic screening. The methods proposed herein, at the throughput enabled by IR-MALDESI, could offer a new avenue for the discovery of novel drugs.http://www.sciencedirect.com/science/article/pii/S2472630325000305IR-MALDESIPhenotypic screeningDrug discoveryCell-based assayGlutaminaseMass spectrometry |
| spellingShingle | Alena N. Joignant Fan Pu Shaun M. McLoughlin James W. Sawicki Andrew J. Radosevich Renze Ma Jon D. Williams Sujatha M. Gopalakrishnan Nathaniel L. Elsen Metabolite fingerprinting by infrared matrix-assisted laser desorption electrospray ionization mass spectrometry SLAS Technology IR-MALDESI Phenotypic screening Drug discovery Cell-based assay Glutaminase Mass spectrometry |
| title | Metabolite fingerprinting by infrared matrix-assisted laser desorption electrospray ionization mass spectrometry |
| title_full | Metabolite fingerprinting by infrared matrix-assisted laser desorption electrospray ionization mass spectrometry |
| title_fullStr | Metabolite fingerprinting by infrared matrix-assisted laser desorption electrospray ionization mass spectrometry |
| title_full_unstemmed | Metabolite fingerprinting by infrared matrix-assisted laser desorption electrospray ionization mass spectrometry |
| title_short | Metabolite fingerprinting by infrared matrix-assisted laser desorption electrospray ionization mass spectrometry |
| title_sort | metabolite fingerprinting by infrared matrix assisted laser desorption electrospray ionization mass spectrometry |
| topic | IR-MALDESI Phenotypic screening Drug discovery Cell-based assay Glutaminase Mass spectrometry |
| url | http://www.sciencedirect.com/science/article/pii/S2472630325000305 |
| work_keys_str_mv | AT alenanjoignant metabolitefingerprintingbyinfraredmatrixassistedlaserdesorptionelectrosprayionizationmassspectrometry AT fanpu metabolitefingerprintingbyinfraredmatrixassistedlaserdesorptionelectrosprayionizationmassspectrometry AT shaunmmcloughlin metabolitefingerprintingbyinfraredmatrixassistedlaserdesorptionelectrosprayionizationmassspectrometry AT jameswsawicki metabolitefingerprintingbyinfraredmatrixassistedlaserdesorptionelectrosprayionizationmassspectrometry AT andrewjradosevich metabolitefingerprintingbyinfraredmatrixassistedlaserdesorptionelectrosprayionizationmassspectrometry AT renzema metabolitefingerprintingbyinfraredmatrixassistedlaserdesorptionelectrosprayionizationmassspectrometry AT jondwilliams metabolitefingerprintingbyinfraredmatrixassistedlaserdesorptionelectrosprayionizationmassspectrometry AT sujathamgopalakrishnan metabolitefingerprintingbyinfraredmatrixassistedlaserdesorptionelectrosprayionizationmassspectrometry AT nathaniellelsen metabolitefingerprintingbyinfraredmatrixassistedlaserdesorptionelectrosprayionizationmassspectrometry |