2010
DOI: 10.1021/ac101024z
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Immunomagnetic Separation and Quantification of Butyrylcholinesterase Nerve Agent Adducts in Human Serum

Abstract: A novel method for extracting butyrylcholinesterase (BuChE) from serum as a means of identifying and measuring nerve agent adducts to human BuChE is presented here. Antibutyrylcholinesterase monoclonal antibodies were conjugated to protein-G ferromagnetic particles and mixed with 500 microL serum samples. The particle-antibody-BuChE product was rinsed and directly digested with pepsin. Native and isotopically enriched nonapeptides corresponding to the pepsin digest products for uninhibited BuChE, and sarin, cy… Show more

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Cited by 122 publications
(169 citation statements)
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“…Both LC and MALDI-TOF MS have been used in the identification of OP exposures from either insecticides or nerve agents (Sun and Lynn, 2007;Thompson et al, 2010;Noort and Fidder, 2006;Li et al, 2008;Sun and Lynn., 2009;Li et al, 2010;Schopfer et al, 2010;Liyasova et al, 2011). In order to remove the limitations of affinity chromatography for biomarker purification, immunomagnetic beads coupled to a specific antiBChE antibody have been used in other studies that use LC-MS/MS to monitor OP exposures (Kim et al, 2010;Sporty et al, 2010;Marsillach et al, 2011;Aryal et al, 2012). The LC-MS/MS analysis of OP exposures has shown to have several advantages over the currently used colorimetric enzyme activity assays: 1) the analysis does not require a separate blood draw for the determination of baseline activity levels; 2) it provides a more accurate measurement of the percentage modification of the active-site serine, and; 3) it also provides some information about the OP that adducted the active-site serine (e.g., a methyl-or ethyl based insecticide).…”
Section: Mass Spectrometry (Ms) Studiesmentioning
confidence: 99%
“…Both LC and MALDI-TOF MS have been used in the identification of OP exposures from either insecticides or nerve agents (Sun and Lynn, 2007;Thompson et al, 2010;Noort and Fidder, 2006;Li et al, 2008;Sun and Lynn., 2009;Li et al, 2010;Schopfer et al, 2010;Liyasova et al, 2011). In order to remove the limitations of affinity chromatography for biomarker purification, immunomagnetic beads coupled to a specific antiBChE antibody have been used in other studies that use LC-MS/MS to monitor OP exposures (Kim et al, 2010;Sporty et al, 2010;Marsillach et al, 2011;Aryal et al, 2012). The LC-MS/MS analysis of OP exposures has shown to have several advantages over the currently used colorimetric enzyme activity assays: 1) the analysis does not require a separate blood draw for the determination of baseline activity levels; 2) it provides a more accurate measurement of the percentage modification of the active-site serine, and; 3) it also provides some information about the OP that adducted the active-site serine (e.g., a methyl-or ethyl based insecticide).…”
Section: Mass Spectrometry (Ms) Studiesmentioning
confidence: 99%
“…9,10 Laboratory-based GC/LC/MS analytical techniques are commonly used and show detection limits in the nanomolar concentration range. However, these techniques are time-consuming, expensive, have to be performed by a highly trained technician, and not suitable for rapid analyses under field conditions, especially in emergency cases.…”
mentioning
confidence: 99%
“…(Carter et al, 2013, Fidder et al, 2002, Knaack et al, 2012, Sporty et al, 2010) When adducted BChE peptides (FGESAGAAS) are fragmented using collision-induced dissociation, β-elimination of the OPNA adduct is observed, converting the adducted serine into a dehydroalanine ( m/z 778.3) (Figure 4). For the MeP-BChE, EtP-BChE, PrP-BChE, and P-BChE peptides, quantitation was based on the transition of each respective [M+H] + precursor ion to the product ion m/z 778.3.…”
Section: Resultsmentioning
confidence: 99%
“…OPNA-BChE adducts have relatively long half-lives (8–12 days), are abundant in serum and plasma, and can easily be extracted from blood since BChE is not membrane-bound. (Knaack et al, 2012, Pantazides et al, 2014, Sporty et al, 2010)…”
Section: Introductionmentioning
confidence: 99%