Multidimensional chromatography coupled with mass spectrometry for target-based screening

Hsieh, Yinliang F.; Gordon, Neal F.; Regnier, Fred E.; Afeyan, Noubar B.; Martin, Stephen A.; Vella, George

doi:10.1007/bf01715634

Cited by 28 publications

(22 citation statements)

References 19 publications

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“…95 Similarly, combinatorial library analysis using various affinity-based MS approaches have been utilized to depict trends in the SAR of the structural determinants of synthetic chemical analogues. [66][67][68][69][70]74,75 …”

Section: The Importance Of Specificitymentioning

confidence: 99%

Strategic Use of Affinity-Based Mass Spectrometry Techniques in the Drug Discovery Process

2001

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Advances in biomolecular mass spectrometry (Bio-MS) have made this technique an invaluable tool for analytical chemists and biochemists alike. The applicability of Bio-MS approaches in drug discovery now encompasses in vitro, cellular, and in vivo pharmacological and clinical applications in an unprecedented expansion of utility. As a result, the role of Bio-MS in pharmaceutical discovery continues to proliferate for both structural and functional characterization of biomolecules. From target characterization to lead optimization, affinity techniques have been used to purify, probe, and enrich analytes of interest. Affinity selection employed prior to MS analysis can "edit" out extraneous noise and enable the researcher to examine only what is important. These affinity-based methods can be used as an alternative strategy when classical biochemical techniques are insufficient in advancing difficult projects. We have applied various affinity techniques in conjunction with mass spectrometry throughout the drug discovery process. This perspective will describe affinity-based mass spectrometry methodologies and related concepts, illustrated with original results.

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Section: The Importance Of Specificitymentioning

confidence: 99%

Strategic Use of Affinity-Based Mass Spectrometry Techniques in the Drug Discovery Process

2001

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“…Following SEC and desalting/complex dissociation using a peptide cartridge, the ligands were eluted into a triple quadrupole mass spectrometer with ESI. An automated multi‐dimensional chromatographic system based on SEC and reversed‐phase separation coupled to ESI was presented by Hsieh et al 64…”

Section: Ms‐based Screening Of Libraries In Solutionmentioning

confidence: 99%

Mass spectrometry and combinatorial chemistry: a short outline

et al. 2001

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The rapid evolution of combinatorial chemistry in recent years has led to a dramatic improvement in synthetic capabilities. The goal is to accelerate the discovery of molecules showing affinity against a target, such as an enzyme or a receptor, through the simultaneous synthesis of a great number of structurally diverse compounds. This is done by generating combinatorial libraries containing as many as hundreds or thousands of compounds. The need to test all these compounds led to the development of high-throughput screening (HTS) techniques, and also high-throughput analytical techniques capable of assessing the occurrence, structure and purity of the products. In order to be applied effectively to the characterization of combinatorial libraries, an analytical technique must be adequately sensitive (to analyse samples which are typically produced in nanomole amounts or less), fast, affordable and easy to automate (to minimize analysis time and operator intervention). Although no method alone can meet all the analytical challenges underlying this task, the recent progress in mass spectrometric (MS) instrumentation renders this technique an essential tool for scientists working in this area. We describe here relevant aspects of the use of MS in combinatorial technologies, such as current methods of characterization, purification and screening of libraries. Some examples from our laboratory deal with the analysis of pooled oligomeric libraries containing n x 324(n = 1, 2) compounds, using both on-line high-performance liquid chromatography/MS with an ion trap mass spectrometer, and direct infusion into a triple quadrupole instrument. In the first approach, MS and product ion MS/MS with automatic selection of the precursor were performed in one run, allowing library confirmation and structural elucidation of unexpected by-products. The second approach used MS scans to characterize the entire library and also precursor ion and neutral loss scans to detect selectively components with given structural characteristics.

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“…A variant of this procedure has been incorporated into an automated screening methodology termed Selectronics ™ and is illustrated in Figure 24. 174 In addition, this general screening procedure has been used in competitive binding experiments to identify library components with optimum binding affinity. 175 Gel-filtration separations are best suited to ligands that have high affinity for the receptor molecule (K d < 1 µM).…”

Section: Ligand Selection By Gel-filtrationmentioning

confidence: 99%

Combinatorial Chemistry Libraries, Analysis of

Kibbey¹

2000

Encyclopedia of Analytical Chemistry

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Many of the analytical techniques applied to the analysis of traditional pharmaceutical compounds are used to characterize combinatorial chemistry libraries. In both cases, the medicinal chemist is primarily concerned with verifying the identity and purity of the compounds synthesized. The identity of a compound usually is confirmed spectroscopically by Fourier transform infrared (FTIR) spectroscopy, mass spectrometry (MS), and/or nuclear magnetic resonance (NMR) spectroscopy, while purity is determined using a chromatographic technique, such as high‐performance liquid chromatography (HPLC). Combinatorial chemistry libraries may contain 100–100 000 compounds present as single species or as mixtures. The need to efficiently characterize such large numbers of compounds has been the primary motivation behind the development of many of the chromatographic–spectroscopic techniques applied to the analysis of combinatorial chemistry libraries. Many of these techniques have utility outside of the realm of normal library characterization. For example, the combination of affinity chromatography and MS has proven useful for screening combinatorial chemistry libraries against specific biological targets. The major impediments to the analysis of combinatorial chemistry libraries have been the challenges of designing analytical methods compatible with the wide chemical diversity of compounds encountered in these libraries, and the unique sample preparation and handling requirements imposed by combinatorial synthesis.

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Multidimensional chromatography coupled with mass spectrometry for target-based screening

Cited by 28 publications

References 19 publications

Strategic Use of Affinity-Based Mass Spectrometry Techniques in the Drug Discovery Process

Strategic Use of Affinity-Based Mass Spectrometry Techniques in the Drug Discovery Process

Mass spectrometry and combinatorial chemistry: a short outline

Combinatorial Chemistry Libraries, Analysis of

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