Quantitative Mass Spectrometry Evaluation of Human Retinol Binding Protein 4 and Related Variants 英文参考文献.docVIP

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Quantitative Mass Spectrometry Evaluation of Human Retinol Binding Protein 4 and Related Variants 英文参考文献.doc

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Quantitative Mass Spectrometry Evaluation of Human Retinol Binding Protein 4 and Related Variants 英文参考文献

QuantitativeMassSpectrometryEvaluationofHuman RetinolBindingProtein4andRelatedVariants UrbanA.Kiernan*,DavidA.Phillips,OlgicaTrenchevska,DobrinNedelkov IntrinsicBioprobes,Inc.,Tempe,Arizona,UnitedStatesofAmerica Abstract Background:RetinolBindingProtein4(RBP4)isanexcitingnewbiomarkerforthedeterminationofinsulinresistanceand type2diabetes.ItisknownthatcirculatingRBP4residesinmultiplevariantswhichmayprovideenhancedclinicalutility, but conventional immunoassay methods are blind to such differences. A Mass Spectrometric immunoassay (MSIA) technology that can quantitate total RBP4 as well as individual isoforms may provide an enhanced analysis for this biomarker. Methods: RBP4 was isolated and detected from 0.5uL of human plasma using MSIA technology, for the simultaneous quantificationanddifferentiationofendogenoushumanRBP4anditsvariants. Results: The linear range of the assay was 7.81–500ug/mL, and the limit of detection and limit of quantification were 3.36ug/mLand6.52ug/mL,respectively. Theintra-assayCVsweredeterminedtobe5.1%andtheinter-assayCVswere 9.6%.ThepercentrecoveryoftheRBP4-MSIArangedfrom95–105%.MethodcomparisonoftheRBP4MSIAvstheImmun DiagnostikELISAyieldedaPassingBablokfitofMSIA =1.056ELISA–3.09,whiletheCusumlinearityp-valuewas.0.1 andthemeanbiasdeterminedbytheAltmanBlandtestwas1.2%. Conclusion:ThenovelRBP4MSIAprovidedafast,accurateandprecisequantitativeproteinmeasurementascomparedto thestandardcommerciallyavailableELISA.Moreover,thismethodalsoallowedforthedetectionofRBP4variantsthatare presentineachsample,whichmayinthefutureprovideanewdimensionintheclinicalutilityofthisbiomarker. Citation:KiernanUA,PhillipsDA,TrenchevskaO,NedelkovD(2011)QuantitativeMassSpectrometryEvaluationofHumanRetinolBindingProtein4andRelated Variants.PLoSONE6(3):e17282.doi:10.1371/journal.pone.0017282 Editor:MassimoFederici,UniversityofTorVergata,Italy ReceivedSeptember7,2010;AcceptedJanuary28,2011;PublishedMarch30,2011 Copyright: ? 2011 Kiernan et al. This is an open-access ar