Affinity Capillary Electrophoresis−Mass Spectrometry for Screening Combinatorial Libraries

Chu, Yen‐Ho; Dunayevskiy, Yuriy M.; Kirby, Daniel P.; Vouros, and Paul; Karger, Barry L.

doi:10.1021/ja960213h

Cited by 168 publications

(115 citation statements)

References 25 publications

Supporting

Mentioning

113

Contrasting

Order By: Relevance

“…Affinity capillary electrophoresis combined with MS was first used for this purpose. 164 This method can simultaneously measure the binding affinities of multiple ligands, generated by combinatorial chemistry, for a target receptor based on mobility shifts on capillary electrophoregrams, and can monitor the MS and MS/MS spectra of every active molecule for structural determination with high sensitivity. Affinity chromatography coupled with MS using ligand, receptor, antibody, or enzyme immobilized affinity beads in place of affinity capillary electrophoresis has been reported for library affinity selection.…”

Section: ·1 High-throughput Analysismentioning

confidence: 99%

Biomedical and Biological Mass Spectrometry

Mano

Goto

2003

ANAL. SCI.

View full text Add to dashboard Cite

By the end of the 20th century, mass spectrometry (MS) had evolved drastically as a major analytical tool in biomedical and biological research. Recent mass spectrometers, designed for ease of handling, came into wide use in the 1990s. Soft ionization methods, which were made practical in the second half of the 1980s, allowed for the sensitive analysis of nonvolatile and thermolabile compounds by MS. Electrospray ionization (ESI) 1,2 and matrix-assisted laser desorption ionization (MALDI) 3 methods, made it possible to ionize large biomolecules and to transfer them to the gas phase, thereby becoming a valuable tool in both protein and peptide analysis [4][5][6] and in the study of drugs and their metabolites [7][8][9][10][11][12] in the human body. Many reviews of MS have recently been published, [13][14][15][16][17][18][19] and in particular, the number of reviews describing biochemical, biological, and biomedical MS have increased dramatically over the last ten years. Initially, there were many commentary reviews on new techniques, such as soft ionization methods and mass analyzers, 13-15 and more recently, review articles have focused on the biological and biomedical applications of MS. 18-21 Above all, two major fields, drug discovery and proteomics, have made marked advances based on MS.Articles in the MEDLINE database were searched, and the results are illustrated in Fig. 1 and Table 1. Over the last 15 years, a total of 79220 MS articles were placed in the database, with a 2.5-fold increase from 1987 to 1999. The number of articles using fast atom bombardment (FAB) as an ionization method remained almost constant during this period. This ionization method is a practical soft ionization procedure, which was applied to the analysis of peptides [22][23][24][25][26][27][28][29][30][31] and endogenous biological substances [32][33][34][35] in the 1980s. Atmospheric pressure chemical ionization (APCI) is a suitable method 36 for the ionization of middle or weakly polar low-molecular-weight molecules, and can be easily coupled to conventional highperformance liquid chromatography (HPLC), used at a flow rate of around 1 mL/min. [37][38][39][40][41][42][43][44][45][46][47][48] However, the number of articles concerning APCI have increased slowly, because most drugs developed recently contain polar groups which can be ionized without difficulty by the ESI technique. Japan This review focuses on biological and biomedical mass spectrometry, and covers a selection of publications in this area included in the MEDLINE database for the period 1987 -2001. Over the last 15 years, biological and biomedical mass spectrometry has progressed out of all recognition. The development of soft ionization methods, such as electrospray ionization and matrix-assisted laser desorption ionization, has mainly contributed to the remarkable progress, because they can easily produce gas-phase ions of large, polar, and thermally labile biomolecules, such as proteins, peptides, nucleic acids and others. The innovations of ionization meth...

show abstract

Section: ·1 High-throughput Analysismentioning

confidence: 99%

Biomedical and Biological Mass Spectrometry

Mano

Goto

2003

ANAL. SCI.

View full text Add to dashboard Cite

show abstract

“…By combining affinity capillary electrophoresis with on-line mass spectrometric detection and identification, affinity constants for multiple compounds can be measured in a single analysis [6]. Recognizing that on-line mass spectrometric detection was helpful for the identification of each ligand, Chu et al [48] extended this approach to include the screening of combinatorial libraries as a means of drug discovery.…”

Section: Affinity Capillary Electrophoresis-mass Spectrometrymentioning

confidence: 99%

“…Also, note that the types of libraries that have been screened using this approach have contained modest numbers of synthetic analogs such as peptides. Libraries exceeding 400 members required preliminary purification using affinity chromatography to reduce the number of compounds [48]. As a result, this approach is probably not ideal for screening large molecularly diverse libraries or natural product extracts.…”

Section: Affinity Capillary Electrophoresis-mass Spectrometrymentioning

confidence: 99%

Analysis and screening of combinatorial libraries using mass spectrometry

Shin

Breemen

2001

Biopharm & Drug Disp

View full text Add to dashboard Cite

Mass spectrometry is a highly selective and high throughput analytical technique that is ideally suited for the identification and purity determination of large numbers of compounds prepared using combinatorial chemistry or for the dereplication of natural products. Compounds may be characterized based on molecular weight, elemental composition and structural features based on fragmentation patterns. When coupled to a separation technique such as highperformance liquid chromatography (HPLC) or capillary electrophoresis, mass spectrometric applications may be expanded to include analysis of complex mixtures. However, the slower speed of the separation step reduces the throughput of the analysis. This review concerns the application of mass spectrometry to the characterization of combinatorial libraries and the screening of library and natural product mixtures. Strategies to enhance the throughput of LC-MS are discussed including fast HPLC and parallel LC-MS. Also, mass spectrometry-based screening methods are described including frontal affinity chromatography-mass spectrometry, gel permeation chromatography LC-MS, direct electrospray mass spectrometry of receptor-ligand complexes, affinity chromatography-mass spectrometry, and pulsed ultrafiltration mass spectrometry.

show abstract

“…Since mass spectrometry provides excellent selectivity, sensitivity, and structural information to facilitate the identification of ligands in complex mixtures, various screening assays have been developed based on multidimensional chromatography mass spectrometry, such as sizeexclusion chromatography LC-MS [6], ultrafiltration LC-MS [7], affinity capillary electrophoresis-MS [8], H/D exchange MALDI MS [9], and FT-ICR MS [10]. For example, we have reported the development of ultrafiltration LC-MS to screen mixtures of compounds for ligands to various receptors, such as estrogen receptors (ER) [11], cyclooxygenase-2 [12] and human serum albumin [13].…”

Section: Introductionmentioning

confidence: 99%

Development of a Screening Assay for Ligands to the Estrogen Receptor Based on Magnetic Microparticles and LC-MS

Choi¹,

Breemen²

2008

CCHTS

View full text Add to dashboard Cite

A high throughput screening assay for the identification of ligands to pharmacologically significant receptors was developed based on magnetic particles containing immobilized receptors followed by liquid chromatography-mass spectrometry (LC-MS). This assay is suitable for the screening of complex mixtures such as botanical extracts. For proof-of-principle, estrogen receptor-α (ER-α) and ER-β were immobilized on magnetic particles functionalized with aldehyde or carboxylic acid groups. Alternatively, biotinylated ER was immobilized onto streptavidin-derivatized magnetic particles. The ER that was immobilized using the streptavidin-biotin chemistry showed higher activity than that immobilized on aldehyde or carboxylic acid functionalized magnetic particles. Immobilized ER was incubated with extracts of Trifolium pratense L. (red clover) or Humulus lupulus L. (hops).As a control for non-specific binding, each botanical extract was incubated with magnetic particles containing no ER. After magnetic separation of the particles containing bound ligands from the unbound components in the extract, the particles were washed, ligands were released using methanol, and then the ligands were identified using LC-MS. The estrogens genistein and daidzein were identified in the red clover extract, and the estrogen 8-prenylnaringenin was identified in the hop extract. These screening results are consistent with those obtained using previous screening approaches.

show abstract

Affinity Capillary Electrophoresis−Mass Spectrometry for Screening Combinatorial Libraries

Cited by 168 publications

References 25 publications

Biomedical and Biological Mass Spectrometry

Biomedical and Biological Mass Spectrometry

Analysis and screening of combinatorial libraries using mass spectrometry

Development of a Screening Assay for Ligands to the Estrogen Receptor Based on Magnetic Microparticles and LC-MS

Contact Info

Product

Resources

About