Evaluation of high performance liquid chromatography/electrospray mass spectrometry with selected ion monitoring for the analysis of large synthetic combinatorial peptide libraries

Lambert, Pierre‐Herve; Bertin, Sophie; Fauchère, Jean‐Luc; Volland, Jean‐Paul

doi:10.1002/(sici)1097-0231(199712)11:18<1971::aid-rcm51>3.0.co;2-8

Cited by 19 publications

(7 citation statements)

References 19 publications

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“…Alternative experimental procedures, such as HPLC/MS, would give a more accurate measure of the extent of reaction and final product distribution, while HPLC/MS/MS would yield qualitative, sequence and structural information 19,. 20 However, such experimental methods were not included in this study, instead, we have focused on the potentials and advantages of high resolution mass separation and exact mass measurement by FTICR‐MS in order to illustrate the influence of monisotopic and carbon‐13 isobars on the identification of components of the dodecapeptide library.…”

Section: Resultsmentioning

confidence: 99%

“…Interfacing high performance liquid chromatography with nuclear magnetic resonance (HPLC/NMR) has been particularly useful in characterizing geometric and isobaric molecules contained in numerous peptide libraries 16–18. However, the more productive high throughput methodologies are designed around HPLC/MS, and HPLC/MS/MS, whereby both qualitative and quantitative results can be achieved 19–23. Fourier transform ion cyclotron resonance mass spectrometry (FTICR‐MS), coupled with electrospray ionization (ESI) has been successfully applied to the characterization of peptide and non‐peptidic combinatorial libraries 24–28.…”

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confidence: 99%

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The significance of monoisotopic and carbon-13 isobars for the identification of a 19-component dodecapeptide library by positive ion electrospray Fourier transform ion cyclotron resonance mass spectrometry

Ramjit¹,

Kruppa²,

Spier³

et al. 2000

Rapid Commun. Mass Spectrom.

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Harnessing the ultra high resolution capabilities of Fourier transform ion cyclotron resonance mass spectrometry (FTICR‐MS) and positive ion electrospray, we have demonstrated the significance and utility of cumulative mass defect high resolution mass separation stable isotope distribution, exact mass measurement and elemental formula as a means of simultaneously identifying 19 components of the dodecapeptide library Ac‐ANKISYQS[X]STE‐NH2. With an instrument resolution of 275 000 (average), isobaric multiplets attributed to monoisotopic and carbon‐13 components of peptides: Ac ∼ SLS ∼ NH2; Ac ∼ SNS ∼ NH2; Ac ∼ SOS ∼ NH2; Ac ∼ SDS ∼ NH2; within the mass window of 1380–1385 Da, and Ac ∼ SQS ∼ NH2; Ac ∼ SKS ∼ NH2; Ac ∼ SES ∼ NH2; Ac ∼ SMS ∼ NH2, within the mass window 1395–1400 Da, were mass resolved, accurately mass measured and identified from the computed molecular formulas. This experimental procedure enabled the separation of monoisotopic and carbon‐13 isobars yielding enhanced selectivity and specificity and serves to illustrate the significance of monoisotopic and carbon‐13 isobars in final product analysis. Chromatographic separation (HPLC) was of limited utility except for monitoring the overall extent of reaction and apparent product distribution. Positive ion electrospray‐FTICR‐MS and fast atom bombardment (FAB) MS were used to assess final product quality and apparent component distribution. Copyright © 2000 John Wiley & Sons, Ltd.

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

The significance of monoisotopic and carbon-13 isobars for the identification of a 19-component dodecapeptide library by positive ion electrospray Fourier transform ion cyclotron resonance mass spectrometry

Ramjit¹,

Kruppa²,

Spier³

et al. 2000

Rapid Commun. Mass Spectrom.

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“…The final compounds were obtained after cleavage and underwent deprotection in 95% TFA in the presence of scavengers, evaporation, suspension in diethylether, centrifugation and finally lyophilization in acetonitrile/water which afforded powders. Each of the resulting 24 sublibraries was analyzed using MS and MS/MS techniques (14,15) providing enough information to ensure the lack of major by‐products and the presence of the expected structures in the mixture. Furthermore, in each sublibrary, individual amino acids were indeed found to be present in roughly equal amounts by two‐dimensional NMR (14).…”

Section: Peptide Library Synthesis and Deconvolutionmentioning

confidence: 99%

From peptide libraries to optimized nonpeptide ligands in the search for S‐farnesyltransferase inhibitors

Henlin

Kucharczyk

Desmet-Beaufort

et al. 2001

The Journal of Peptide Research

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A complete 331,776-member library of tetrapeptides made of 24 amino acid building blocks was synthesized robotically on solid phase and subjected to a deconvolution based on the inhibitory potency of the sublibraries in a HPLC assay of the S-farnesyltransferase activity in vitro. One of the non-natural peptide and noncysteine-containing leads Nip-Trp-Phe-His (Nip=p-nitrophenyl-L-alanine) was optimized chemically to give a proteolytically stable pseudopeptide with a 200-fold potency compared with the original lead. The final compound was converted to the C-terminal ethyl ester: p-F-C6H4-CO(CH2)2-CO-Bta-D-Phepsi[CH2NH]His-OEt (Bta = benzothienyl-L-alanine) and shown to behave as a prodrug which was hydrolyzed back to the C-terminal acid following cell penetration. The method confirmed that several structurally original leads can be discovered in large libraries when deconvolution relies upon a highly specific assay and that these leads can be optimized by chemical modification to impart the final compound the desired pharmacological and pharmacokinetic properties.

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“…Ionization yields for individual peptides present in mixtures with ESIMS, however, have been reported to be nearly identical. 98 Lambert et al 99 reported that a single peptide possessing a unique mass may be identified from a library containing up to 100 000 peptides using a combination of LC/ESIMS with selected ion monitoring. The feasibility of characterizing peptide mixtures by MS has been explored mathematically using computer simulations of mass spectra for peptides containing 2 -7 amino acids.…”

Section: Electrospray Ionization Mass Spectrometrymentioning

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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Evaluation of high performance liquid chromatography/electrospray mass spectrometry with selected ion monitoring for the analysis of large synthetic combinatorial peptide libraries

Cited by 19 publications

References 19 publications

The significance of monoisotopic and carbon-13 isobars for the identification of a 19-component dodecapeptide library by positive ion electrospray Fourier transform ion cyclotron resonance mass spectrometry

The significance of monoisotopic and carbon-13 isobars for the identification of a 19-component dodecapeptide library by positive ion electrospray Fourier transform ion cyclotron resonance mass spectrometry

From peptide libraries to optimized nonpeptide ligands in the search for S‐farnesyltransferase inhibitors

Combinatorial Chemistry Libraries, Analysis of

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