کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5136551 | 1494016 | 2017 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Plasma PCSK9 measurement by liquid chromatography-Tandem mass spectrometry and comparison with conventional ELISA
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کلمات کلیدی
DTTdithiotreitolPCSK9LDL-CSDCLC–MS/MS - LC-MS / MSinternal standard - استاندارد داخلیSolid-phase extraction - استخراج فاز جامدstandard deviation - انحراف معیارAmmonium bicarbonate - بی کربنات آمونیومEnzyme-linked immunosorbent assay - تست الیزاELISA - تست الیزاSodium deoxycholate - سدیم دگزیکسولاتCoefficient of Variation - ضریب تغییرLow-density lipoprotein - لیپوپروتئین کم چگالی یا الدیال LDL - لیپوپروتئین کم چگالی(کلسترول بد)Proprotein convertase subtilisin/kexin type 9 - پروتیکین تبدیل اساتید سوتیلیسین / نوع ککسین 9LDL cholesterol - کلسترول LDLiodoacetamide - یووداکتامید
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The combination of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and trypsin proteolysis is an effective tool for accurate quantitation of multiple proteins in a single run. However, expensive samples pre-treatment as immunoenrichment are often required to analyze low abundant proteins. Plasma proprotein convertase subtilisin/kexin type 9 (PCSK9), a circulating regulator of low-density lipoprotein metabolism, was studied as an example of a low abundant plasma protein. We investigated post-proteolysis solid-phase extraction (SPE) as an alternative strategy to improve its detection. After optimization of pretreatment, including denaturation, reduction, alkylation, tryptic digestion and selective SPE concentration, 91 ± 7% of PCSK9 was recovered from human plasma samples and coefficients of variation were less than 13.2% with a lower limit of quantification of 37.5 ng/ml. This LC-MS/MS method was compared with standard enzyme-linked immunosorbent assay in 30 human plasma samples with a broad range of PCSK9 concentrations. Both methods were significantly correlated (r = 0.936, p < 0.001) with less than 7% of the values out of the 95% confidence interval and similar concentrations were measured using either LC-MS/MS or ELISA methods (514.2 ± 217.2 vs. 504.2 ± 231.0 ng/ml, respectively- p = NS). This method involving SPE is an effective measurement tool for low abundant plasma protein analysis that could be easily included in multiplexed assays.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography B - Volumes 1044â1045, 15 February 2017, Pages 24-29
Journal: Journal of Chromatography B - Volumes 1044â1045, 15 February 2017, Pages 24-29
نویسندگان
Mikaël Croyal, Fanta Fall, Michel Krempf, Aurélie Thédrez, Khadija Ouguerram, Véronique Ferchaud-Roucher, Audrey Aguesse, Stéphanie Billon-Crossouard, Pedro Mata, Rodrigo Alonso, Gilles Lambert, Estelle Nobécourt,