A high-throughput MALDI-TOF MS biochemical screen for small molecule inhibitors of the antigen aminopeptidase ERAP1
MALDI-TOF MS is a powerful analytical technique that provides a fast and label-free readout for in vitro assays in the high-throughput screening (HTS) environment. Here, we describe the development of a novel, HTS compatible, MALDI-TOF MS-based drug discovery assay for the endoplasmic reticulum amin...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-01-01
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| Series: | SLAS Discovery |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S247255522213707X |
| _version_ | 1797944097058586624 |
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| author | Leonie Müller Amy K Burton Chloe L Tayler James E Rowedder Jonathan P Hutchinson Simon Peace Julie M Quayle Melanie V Leveridge Roland S Annan Matthias Trost Rachel E Peltier-Heap Maria Emilia Dueñas |
| author_facet | Leonie Müller Amy K Burton Chloe L Tayler James E Rowedder Jonathan P Hutchinson Simon Peace Julie M Quayle Melanie V Leveridge Roland S Annan Matthias Trost Rachel E Peltier-Heap Maria Emilia Dueñas |
| author_sort | Leonie Müller |
| collection | DOAJ |
| description | MALDI-TOF MS is a powerful analytical technique that provides a fast and label-free readout for in vitro assays in the high-throughput screening (HTS) environment. Here, we describe the development of a novel, HTS compatible, MALDI-TOF MS-based drug discovery assay for the endoplasmic reticulum aminopeptidase 1 (ERAP1), an important target in immuno-oncology and auto-immune diseases. A MALDI-TOF MS assay was developed beginning with an already established ERAP1 RapidFire MS (RF MS) assay, where the peptide YTAFTIPSI is trimmed into the product TAFTIPSI. We noted low ionisation efficiency of these peptides in MALDI-TOF MS and hence incorporated arginine residues into the peptide sequences to improve ionisation. The optimal assay conditions were established with these new basic assay peptides on the MALDI-TOF MS platform and validated with known ERAP1 inhibitors. Assay stability, reproducibility and robustness was demonstrated on the MALDI-TOF MS platform. From a set of 699 confirmed ERAP1 binders, identified in a prior affinity selection mass spectrometry (ASMS) screen, active compounds were determined at single concentration and in a dose-response format with the new MALDI-TOF MS setup. Furthermore, to allow for platform performance comparison, the same compound set was tested on the established RF MS setup, as the new basic peptides showed fragmentation in ESI-MS. The two platforms showed a comparable performance, but the MALDI-TOF MS platform had several advantages, such as shorter sample cycle times, reduced reagent consumption, and a lower tight-binding limit. |
| first_indexed | 2024-04-10T20:34:14Z |
| format | Article |
| id | doaj.art-c0f8fd18f1724d62b41a5843c6740ea7 |
| institution | Directory Open Access Journal |
| issn | 2472-5552 |
| language | English |
| last_indexed | 2024-04-10T20:34:14Z |
| publishDate | 2023-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | SLAS Discovery |
| spelling | doaj.art-c0f8fd18f1724d62b41a5843c6740ea72023-01-25T04:15:56ZengElsevierSLAS Discovery2472-55522023-01-01281311A high-throughput MALDI-TOF MS biochemical screen for small molecule inhibitors of the antigen aminopeptidase ERAP1Leonie Müller0Amy K Burton1Chloe L Tayler2James E Rowedder3Jonathan P Hutchinson4Simon Peace5Julie M Quayle6Melanie V Leveridge7Roland S Annan8Matthias Trost9Rachel E Peltier-Heap10Maria Emilia Dueñas11Newcastle University, Faculty of Medical Sciences, Biosciences Institute, Framlington Place, Newcastle Upon Tyne NE2 4HH, United KingdomGSK, Discovery Analytical, Gunnels Wood Rd, Stevenage SG1 2NY, United KingdomGSK, Discovery Analytical, Gunnels Wood Rd, Stevenage SG1 2NY, United KingdomGSK, Screening, Profiling and Mechanistic Biology, Gunnels Wood Rd, Stevenage SG1 2NY, United KingdomGSK, Screening, Profiling and Mechanistic Biology, Gunnels Wood Rd, Stevenage SG1 2NY, United KingdomGSK, Medicinal Chemistry, Gunnels Wood Rd, Stevenage SG1 2NY, United KingdomGSK, Discovery Analytical, Gunnels Wood Rd, Stevenage SG1 2NY, United KingdomGSK, Screening, Profiling and Mechanistic Biology, Gunnels Wood Rd, Stevenage SG1 2NY, United KingdomGSK, Discovery Analytical, Gunnels Wood Rd, Stevenage SG1 2NY, United KingdomNewcastle University, Faculty of Medical Sciences, Biosciences Institute, Framlington Place, Newcastle Upon Tyne NE2 4HH, United Kingdom; Corresponding authors at: Newcastle University, Faculty of Medical Sciences, Biosciences Institute, Framlington Place, Newcastle Upon Tyne NE2 4HH, United KingdomGSK, Discovery Analytical, Gunnels Wood Rd, Stevenage SG1 2NY, United Kingdom; Corresponding author at: GSK, Discovery Analytical, Gunnels Wood Rd, Stevenage SG1 2NY, United KingdomNewcastle University, Faculty of Medical Sciences, Biosciences Institute, Framlington Place, Newcastle Upon Tyne NE2 4HH, United Kingdom; Corresponding authors at: Newcastle University, Faculty of Medical Sciences, Biosciences Institute, Framlington Place, Newcastle Upon Tyne NE2 4HH, United KingdomMALDI-TOF MS is a powerful analytical technique that provides a fast and label-free readout for in vitro assays in the high-throughput screening (HTS) environment. Here, we describe the development of a novel, HTS compatible, MALDI-TOF MS-based drug discovery assay for the endoplasmic reticulum aminopeptidase 1 (ERAP1), an important target in immuno-oncology and auto-immune diseases. A MALDI-TOF MS assay was developed beginning with an already established ERAP1 RapidFire MS (RF MS) assay, where the peptide YTAFTIPSI is trimmed into the product TAFTIPSI. We noted low ionisation efficiency of these peptides in MALDI-TOF MS and hence incorporated arginine residues into the peptide sequences to improve ionisation. The optimal assay conditions were established with these new basic assay peptides on the MALDI-TOF MS platform and validated with known ERAP1 inhibitors. Assay stability, reproducibility and robustness was demonstrated on the MALDI-TOF MS platform. From a set of 699 confirmed ERAP1 binders, identified in a prior affinity selection mass spectrometry (ASMS) screen, active compounds were determined at single concentration and in a dose-response format with the new MALDI-TOF MS setup. Furthermore, to allow for platform performance comparison, the same compound set was tested on the established RF MS setup, as the new basic peptides showed fragmentation in ESI-MS. The two platforms showed a comparable performance, but the MALDI-TOF MS platform had several advantages, such as shorter sample cycle times, reduced reagent consumption, and a lower tight-binding limit.http://www.sciencedirect.com/science/article/pii/S247255522213707XMALDI-TOF MSHigh-throughput screeningDrug discoveryMass spectrometryEndoplasmic reticulum aminopeptidase 1Major histocompatibility complex |
| spellingShingle | Leonie Müller Amy K Burton Chloe L Tayler James E Rowedder Jonathan P Hutchinson Simon Peace Julie M Quayle Melanie V Leveridge Roland S Annan Matthias Trost Rachel E Peltier-Heap Maria Emilia Dueñas A high-throughput MALDI-TOF MS biochemical screen for small molecule inhibitors of the antigen aminopeptidase ERAP1 SLAS Discovery MALDI-TOF MS High-throughput screening Drug discovery Mass spectrometry Endoplasmic reticulum aminopeptidase 1 Major histocompatibility complex |
| title | A high-throughput MALDI-TOF MS biochemical screen for small molecule inhibitors of the antigen aminopeptidase ERAP1 |
| title_full | A high-throughput MALDI-TOF MS biochemical screen for small molecule inhibitors of the antigen aminopeptidase ERAP1 |
| title_fullStr | A high-throughput MALDI-TOF MS biochemical screen for small molecule inhibitors of the antigen aminopeptidase ERAP1 |
| title_full_unstemmed | A high-throughput MALDI-TOF MS biochemical screen for small molecule inhibitors of the antigen aminopeptidase ERAP1 |
| title_short | A high-throughput MALDI-TOF MS biochemical screen for small molecule inhibitors of the antigen aminopeptidase ERAP1 |
| title_sort | high throughput maldi tof ms biochemical screen for small molecule inhibitors of the antigen aminopeptidase erap1 |
| topic | MALDI-TOF MS High-throughput screening Drug discovery Mass spectrometry Endoplasmic reticulum aminopeptidase 1 Major histocompatibility complex |
| url | http://www.sciencedirect.com/science/article/pii/S247255522213707X |
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