Pulsed Ultrafiltration Mass Spectrometry: A New Method for Screening Combinatorial Libraries
| Autor: | Duane L. Venton, Yongzhong Zhao, Chao Ran Huang, Dejan Nikolic, RB van Breemen, Charles P. Woodbury |
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| Rok vydání: | 1997 |
| Předmět: |
Electrospray
Adenosine Deaminase Ultrafiltration Ligands Mass spectrometry Online Systems Mass Spectrometry Analytical Chemistry Furosemide medicine Animals Humans Enzyme Inhibitors Serum Albumin Chromatography Adenine binding Drug discovery Elution Chemistry Adenine Methanol Human serum albumin Salicylates Intestines Molecular Weight Dissociation constant Thyroxine Ultrafiltration (renal) Drug Design Pulsatile Flow Cattle Warfarin Salicylic Acid medicine.drug |
| Zdroj: | Analytical Chemistry. 69:2159-2164 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/ac970132j |
| Popis: | In response to the need for rapid screening of combinatorial libraries to identify new lead compounds during drug discovery, we have developed an on-line combination of ultrafiltration and electrospray mass spectrometry, called pulsed ultrafiltration mass spectrometry, which facilitates the identification of solution-phase ligands in library mixtures that bind to solution-phase receptors. After ligands contained in a library mixture were bound to a macromolecular receptor, e.g., human serum albumin or calf intestine adenosine deaminase, the ligand-receptor complexes were purified by ultrafiltration and then dissociated using methanol to elute the ligands into the electrospray mass spectrometer for detection. Ligands with dissociation constants in the micromolar to nanomolar range were successfully bound, released, and detected using this method, including warfarin, salicylate, furosemide, and thyroxine binding to human serum albumin, and erythro-9-(2-hydroxy-3-nonyl)adenine binding to calf intestine adenosine deaminase. Repetitive bind- and-release experiments demonstrated that the receptor could be reused. Thus, pulsed ultrafiltration mass spectrometry was shown to provide a simple and powerful new method for the screening of combinatorial libraries in support of new drug discovery. |
| Databáze: | OpenAIRE |
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