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High-throughput fragment screening by affinity LC-MS
- Source :
- Journal of biomolecular screening. 18(2)
- Publication Year :
- 2012
-
Abstract
- Fragment screening, an emerging approach for hit finding in drug discovery, has recently been proven effective by its first approved drug, vemurafenib, for cancer treatment. Techniques such as nuclear magnetic resonance, surface plasmon resonance, and isothemal titration calorimetry, with their own pros and cons, have been employed for screening fragment libraries. As an alternative approach, screening based on high-performance liquid chromatography separation has been developed. In this work, we present weak affinity LC/MS as a method to screen fragments under high-throughput conditions. Affinity-based capillary columns with immobilized thrombin were used to screen a collection of 590 compounds from a fragment library. The collection was divided into 11 mixtures (each containing 35 to 65 fragments) and screened by MS detection. The primary screening was performed in4 h (corresponding to3500 fragments per day). Thirty hits were defined, which subsequently entered a secondary screening using an active site-blocked thrombin column for confirmation of specificity. One hit showed selective binding to thrombin with an estimated dissociation constant (K (D)) in the 0.1 mM range. This study shows that affinity LC/MS is characterized by high throughput, ease of operation, and low consumption of target and fragments, and therefore it promises to be a valuable method for fragment screening.
- Subjects :
- Drug Evaluation, Preclinical
Computational biology
Pharmacology
Biochemistry
Approved drug
Mass Spectrometry
Analytical Chemistry
Small Molecule Libraries
Fragment (logic)
Liquid chromatography–mass spectrometry
medicine
Humans
Vemurafenib
Drug discovery
Chemistry
Weak affinity chromatography
Thrombin
Cancer treatment
High-Throughput Screening Assays
Molecular Docking Simulation
Kinetics
Molecular Medicine
Biotechnology
medicine.drug
Chromatography, Liquid
Protein Binding
Subjects
Details
- ISSN :
- 1552454X
- Volume :
- 18
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Journal of biomolecular screening
- Accession number :
- edsair.doi.dedup.....6a3673da877247472ccdccd639773df7