Automated MALDI Target Preparation Concept: Providing Ultra-High-Throughput Mass Spectrometry–Based Screening for Drug Discovery
| Autor: | Martin Winter, Carola Kleiner, Robert Ries, Daniel Bischoff, Frank H. Büttner, Tom Bretschneider, Andreas H. Luippold |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Research areas Drug discovery business.industry Computer science High-throughput screening Drug Evaluation Preclinical Ms analysis Mass spectrometry 01 natural sciences Mass spectrometric Automation High-Throughput Screening Assays 0104 chemical sciences Computer Science Applications 010404 medicinal & biomolecular chemistry 03 medical and health sciences Medical Laboratory Technology 030104 developmental biology Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Embedded system Drug Discovery Direct analysis business |
| Zdroj: | SLAS Technology. 24:209-221 |
| ISSN: | 2472-6303 |
| DOI: | 10.1177/2472630318791981 |
| Popis: | Label-free, mass spectrometric (MS) deciphering of enzymatic reactions by direct analysis of substrate-to-product conversion provides the next step toward more physiological relevant assays within drug discovery campaigns. Reduced risk of suffering from compound interference combined with diminished necessity for tailored signal mediators emphasizes the valuable role of label-free readouts. However, MS-based detection has not hitherto met high-throughput screening (HTS) requirements because of the lack of HTS-compatible sample introduction. In the present study, we report on a fully automated liquid-handling concept built in-house to concatenate biochemical assays with matrix-assisted laser desorption/ionization time-of-flight closing this technological gap. The integrated reformatting from 384- to 1536-well format enables cycle times of 0.6 s/sample for automated spotting and 0.4 s/sample for MS analysis, matching the requirements of HTS compatibility. In-depth examination of spotting quality, quantification accuracy, and instrument robustness together with the implementation of a protein tyrosine phosphatase 1B (PTP1B) inhibitor screening (4896 compounds) demonstrate the potential of the heavily inquired HTS integration of the label-free MS readout. Overall, the presented data demonstrate that the introduced automation concept makes label-free MS-based readouts accessible for HTS within drug discovery campaigns but also in other research areas requiring ultrafast MS-based detection. |
| Databáze: | OpenAIRE |
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