Interlaboratory reproducibility of selective reaction monitoring assays using multiple upfront analyte enrichment strategies.
Interlaboratory reproducibility of selective reaction monitoring assays using multiple upfront analyte enrichment strategies.
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DOI：
10.1021/pr300014s
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发表时间：
2012-08-03
影响因子：
4.4
通讯作者：
Lopez MF
中科院分区：
文献类型：
--
作者：
Prakash A;Rezai T;Krastins B;Sarracino D;Athanas M;Russo P;Zhang H;Tian Y;Li Y;Kulasingam V;Drabovich A;Smith CR;Batruch I;Oran PE;Fredolini C;Luchini A;Liotta L;Petricoin E;Diamandis EP;Chan DW;Nelson R;Lopez MF
在过去的几年中，质谱分析已经成为一种补充和潜在取代常规临床核心实验室中标准免疫测定的技术。将质谱法应用于蛋白质和肽测量可以提供包括高灵敏度、多重分析物的能力以及在氨基酸序列水平上的高特异性的优点。在我们之前的研究中，我们证明了当应用标准化标准操作规程（SOP）测量复杂样品中的合成肽时，质谱-选择性反应监测（MS-SRM）测定具有良好的重现性，因为与免疫测定相比，缺乏重现性一直是临床实验室中使用质谱仪的常见批评。此外，基于SRM的蛋白质测定的一个重要警告是，许多低丰度分析物在用MS检测之前需要某种类型的富集。这增加了程序的复杂性，并且增加了不可重现性的可能性，特别是在不同实验室和不同操作员之间。本研究的目的是检测SRM测定法的实验室间重现性，采用各种前期富集策略和不同类型的临床样本（代表常规临床实验室中常见的真实体液）。使用不同的液相色谱-SRM（LC-SRM）平台和先前开发的SOP，在四个不同的实验室中测试了三种不同的先前发表的低丰度分析物富集策略和一种高丰度分析物非富集策略。结果表明，这些测定法确实具有重现性，在真实的世界样本中，所有四个实验室中重要临床蛋白质测量的变异系数小于30%。
Over the past few years, mass spectrometry has emerged as a technology to complement and potentially replace standard immunoassays in routine clinical core laboratories. Application of mass spectrometry to protein and peptide measurement can provide advantages including high sensitivity, the ability to multiplex analytes, and high specificity at the amino acid sequence level. In our previous study, we demonstrated excellent reproducibility of mass spectrometry-selective reaction monitoring (MS-SRM) assays when applying standardized standard operating procedures (SOPs) to measure synthetic peptides in a complex sample, as lack of reproducibility has been a frequent criticism leveled at the use of mass spectrometers in the clinical laboratory compared to immunoassays. Furthermore, an important caveat of SRM-based assays for proteins is that many low-abundance analytes require some type of enrichment before detection with MS. This adds a level of complexity to the procedure and the potential for irreproducibility increases, especially across different laboratories with different operators. The purpose of this study was to test the interlaboratory reproducibility of SRM assays with various upfront enrichment strategies and different types of clinical samples (representing real-world body fluids commonly encountered in routine clinical laboratories). Three different, previously published enrichment strategies for low-abundance analytes and a no-enrichment strategy for high-abundance analytes were tested across four different laboratories using different liquid chromatography-SRM (LC-SRM) platforms and previously developed SOPs. The results demonstrated that these assays were indeed reproducible with coefficients of variation of less than 30% for the measurement of important clinical proteins across all four laboratories in real world samples.
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影响因子：
5.3
作者：
Kote-Jarai Z;Amin Al Olama A;Leongamornlert D;Tymrakiewicz M;Saunders E;Guy M;Giles GG;Severi G;Southey M;Hopper JL;Sit KC;Harris JM;Batra J;Spurdle AB;Clements JA;Hamdy F;Neal D;Donovan J;Muir K;Pharoah PD;Chanock SJ;Brown N;Benlloch S;Castro E;Mahmud N;O'Brien L;Hall A;Sawyer E;Wilkinson R;Easton DF;Eeles RA
通讯作者：
Eeles RA
影响因子：
5
作者：
Emami, Nashmil;Diamandis, Eleftherios P.
通讯作者：
Diamandis, Eleftherios P.
影响因子：
9.3
作者：
Agger SA;Marney LC;Hoofnagle AN
通讯作者：
Hoofnagle AN
DOI：
10.3343/kjlm.2011.31.2.61
发表时间：
2011-04
期刊：
The Korean journal of laboratory medicine
影响因子：
--
作者：
Boja ES;Rodriguez H
通讯作者：
Rodriguez H
影响因子：
6.6
作者：
Liu, AY;Zhang, H;Diamond, DL
通讯作者：
Diamond, DL