Attomole quantitation of protein separations with accelerator mass spectrometry

Vogel, John S.; Grant, Patrick G.; Buchholz, Bruce A.; Dingley, Karen H.; Turteltaub, Kenneth W.

doi:10.1002/1522-2683(200106)22:10<2037::aid-elps2037>3.0.co;2-8

Cited by 22 publications

(20 citation statements)

References 33 publications

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“…We have utilized this technique to develop a powerful strategy for identification of target proteins of isotopically labeled xenobiotics in vivo, combining AMS with high resolution two-dimensional gel electrophoresis, Triton-acid-urea gels, and HPLC to separate complex protein mixtures and biological mass spectrometry (matrixassisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) and LC-ESI-MS) to identify polypeptides bearing 14 C modifications. In addition to confirming previous reports of hemoglobin as a target for modification by benzene (30,31), we also report the identification of the core histones as targets for [ 14 C]benzene metabolites detected at attomole levels in microgram amounts of mouse bone marrow protein.…”

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confidence: 82%

“…Accelerator mass spectrometry (AMS) 1 provides an extremely sensitive method for the detection of low level modifications of proteins (30). We have utilized this technique to develop a powerful strategy for identification of target proteins of isotopically labeled xenobiotics in vivo, combining AMS with high resolution two-dimensional gel electrophoresis, Triton-acid-urea gels, and HPLC to separate complex protein mixtures and biological mass spectrometry (matrixassisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) and LC-ESI-MS) to identify polypeptides bearing 14 C modifications.…”

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confidence: 99%

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Attomole Detection of in Vivo Protein Targets of Benzene in Mice

Williams

Carver

Miranda

et al. 2002

Molecular & Cellular Proteomics

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Modified proteins were detected in liver and bone marrow of mice following treatment with [ 14 C]benzene. Stained sections were excised from one-dimensional and twodimensional gels and converted to graphite to enable 14 C/ 13 C ratios to be measured by accelerator mass spectrometry. Protein adducts of benzene or its metabolites were indicated by elevated levels of 14 C. A number of proteins were identified by in-gel proteolysis and conventional mass spectrometric methods with the low molecular weight proteins identified including hemoglobin and several histones. The incorporation of 14 C was largely proportional to the density of gel staining, giving little evidence that these proteins were specific targets for selective labeling. This was also true for individual histones subfractionated with Triton-acid-urea gels. A representative histone, H4, was isolated and digested with endopeptidase Asp-N, and the resulting peptides were separated by high performance liquid chromatography. 14 C levels in collected fractions were determined, and the peptides were identified by conventional mass spectrometry. The modifications were distributed throughout the protein, and no particular amino acids or groups of amino acids were identified as selective targets. Thus chemical attack by one or more benzene metabolites upon histones was identified and confirmed, but the resulting modifications appeared to be largely nonspecific. This implies high reactivity toward proteins, enabling such attack to occur at multiple sites within multiple targets.

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confidence: 82%

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confidence: 99%

Attomole Detection of in Vivo Protein Targets of Benzene in Mice

Williams

Carver

Miranda

et al. 2002

Molecular & Cellular Proteomics

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“…Aliquots were obtained from the defrosted plasma samples and placed in quartz combustion tubes. The exact sample mass in this process is not important, since AMS measures an isotope ratio that is converted to equivalent amounts of drug per milliliter or milligram of sample using the known isotopic content of the labeled compound and the average or individual carbon content of each sample (Vogel et al, 2001). Reduced samples were pressed into aluminum holders, mounted in a wheel containing up to 58 samples of which 6 represented standards with well defined isotopic compositions, and analyzed by the 1-mV AMS spectrometer at Lawrence Livermore National Laboratory (Ognibene et al, 2002).…”

Section: Lc-ms/ms Of Compound a In Plasmamentioning

confidence: 95%

Evaluation of Microdosing Strategies for Studies in Preclinical Drug Development: Demonstration of Linear Pharmacokinetics in Dogs of a Nucleoside Analog Over a 50-Fold Dose Range

Sandhu¹,

Vogel²,

Rose³

et al. 2004

Drug Metab Dispos

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ABSTRACT:The technique of accelerator mass spectrometry (AMS) was validated successfully and used to study the pharmacokinetics and disposition in dogs of a preclinical drug candidate (7-deaza-2-Cmethyl-adenosine; Compound A), after oral and intravenous administration. The primary objective of this study was to examine whether Compound A displayed linear kinetics across subpharmacological (microdose) and pharmacological dose ranges in an animal model, before initiation of a human microdose study. The AMS-derived disposition properties of Compound A were comparable to data obtained via conventional techniques such as liquid chromatography-tandem mass spectrometry and liquid scintillation counting analyses. Compound A displayed multiphasic kinetics and exhibited low plasma clearance (5.8 ml/min/kg), a long terminal elimination half-life (17.5 h), and high oral bioavailability (103%). Currently, there are no published comparisons of the kinetics of a pharmaceutical compound at pharmacological versus subpharmacological doses using microdosing strategies. The present study thus provides the first description of the full pharmacokinetic profile of a drug candidate assessed under these two dosing regimens. The data demonstrated that the pharmacokinetic properties of Compound A following dosing at 0.02 mg/kg were similar to those at 1 mg/kg, indicating that in the case of Compound A, the pharmacokinetics in the dog appear to be linear across this 50-fold dose range. Moreover, the exceptional sensitivity of AMS provided a pharmacokinetic profile of Compound A, even after a microdose, which revealed aspects of the disposition of this agent that were inaccessible by conventional techniques.The applications of accelerator mass spectrometry (AMS) are broad-ranging and vary from studying environmental and ecological issues such as the isotopic composition of the atmosphere, soil, and water (Hughen et al., 2000;Beck et al., 2001;Keith-Roach et al., 2001;Mironov et al., 2002), to archaeology and volcanology (Stafford et al., 1984;Vogel et al., 1990;Smith et al., 1999), to its use as a bioanalytical tool for nutritional research (Buchholz et al., 1999; Deuker et al., 2000;Weaver and Liebman, 2002). Biomedical applications of AMS and its use in the arena of pharmaceutical research also have been detailed in review articles (Barker and Garner, 1999;Garner, 2000;Turteltaub and Vogel, 2000). To date, most studies on the metabolism and disposition of xenobiotics by AMS have focused on the binding of carcinogens to DNA and proteins (Turteltaub et al., 1990Frantz et al., 1995;Dingley et al., 1999;Li et al., 2003). These studies have demonstrated a linear relationship between dose and DNA adduct formation. Applications of AMS to the field of pharmaceutical sciences have been limited to only a few studies (Kaye et al., 1997;Young et al., 2001;Garner et al., 2002). However, the pharmaceutical industry is becoming increasingly aware of the potential benefits that may accrue from the ultra-high sensitivity afforded by AMS in terms of evalu...

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“…The development of the MWI was driven by the need for basic analytical tools for biomedical studies with a focus on metabolites, enzyme dynamics, elucidation and quantification of biochemistry pathways, and developing data-sets that are appropriate to test flux balance models [e.g., Bucholz et al, 1999;Miyashita et al, 2001;Stewart et al, 2010;Vogel et al, 2001]. The MWI is based on the design of Sessions et al, [2005] The design and initial optimization of the MWI focused on tracer-based applications in the biological sciences where 1-2µl drops of HPLC separated analytes are, via a micro-capillary needle, placed on the wire, dried, and converted to CO 2 .…”

Section: High-performance Liquid Chromatography -Moving Wire Interfacmentioning

confidence: 99%

GAS-HYBRID SOURCE 14C-AMS:Application to terrestrial ecosystem, biological, and environmental research

Guilderson¹

2012

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Attomole quantitation of protein separations with accelerator mass spectrometry

Cited by 22 publications

References 33 publications

Attomole Detection of in Vivo Protein Targets of Benzene in Mice

Attomole Detection of in Vivo Protein Targets of Benzene in Mice

Evaluation of Microdosing Strategies for Studies in Preclinical Drug Development: Demonstration of Linear Pharmacokinetics in Dogs of a Nucleoside Analog Over a 50-Fold Dose Range

GAS-HYBRID SOURCE 14C-AMS:Application to terrestrial ecosystem, biological, and environmental research

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