Hoofnagle Andrew N, Whiteaker Jeffrey R, Carr Steven A, Kuhn Eric, Liu Tao, Massoni Sam A, Thomas Stefani N, Townsend R Reid, Zimmerman Lisa J, Boja Emily, Chen Jing, Crimmins Daniel L, Davies Sherri R, Gao Yuqian, Hiltke Tara R, Ketchum Karen A, Kinsinger Christopher R, Mesri Mehdi, Meyer Matthew R, Qian Wei-Jun, Schoenherr Regine M, Scott Mitchell G, Shi Tujin, Whiteley Gordon R, Wrobel John A, Wu Chaochao, Ackermann Brad L, Aebersold Ruedi, Barnidge David R, Bunk David M, Clarke Nigel, Fishman Jordan B, Grant Russ P, Kusebauch Ulrike, Kushnir Mark M, Lowenthal Mark S, Moritz Robert L, Neubert Hendrik, Patterson Scott D, Rockwood Alan L, Rogers John, Singh Ravinder J, Van Eyk Jennifer E, Wong Steven H, Zhang Shucha, Chan Daniel W, Chen Xian, Ellis Matthew J, Liebler Daniel C, Rodland Karin D, Rodriguez Henry, Smith Richard D, Zhang Zhen, Zhang Hui, Paulovich Amanda G
University of Washington, Seattle, WA;
Fred Hutchinson Cancer Research Center, Seattle, WA;
Clin Chem. 2016 Jan;62(1):48-69. doi: 10.1373/clinchem.2015.250563.
For many years, basic and clinical researchers have taken advantage of the analytical sensitivity and specificity afforded by mass spectrometry in the measurement of proteins. Clinical laboratories are now beginning to deploy these work flows as well. For assays that use proteolysis to generate peptides for protein quantification and characterization, synthetic stable isotope-labeled internal standard peptides are of central importance. No general recommendations are currently available surrounding the use of peptides in protein mass spectrometric assays.
The Clinical Proteomic Tumor Analysis Consortium of the National Cancer Institute has collaborated with clinical laboratorians, peptide manufacturers, metrologists, representatives of the pharmaceutical industry, and other professionals to develop a consensus set of recommendations for peptide procurement, characterization, storage, and handling, as well as approaches to the interpretation of the data generated by mass spectrometric protein assays. Additionally, the importance of carefully characterized reference materials-in particular, peptide standards for the improved concordance of amino acid analysis methods across the industry-is highlighted. The alignment of practices around the use of peptides and the transparency of sample preparation protocols should allow for the harmonization of peptide and protein quantification in research and clinical care.
多年来，基础研究人员和临床研究人员一直在利用质谱技术在蛋白质测量方面所具有的分析灵敏度和特异性。临床实验室现在也开始采用这些工作流程。对于使用蛋白水解来生成用于蛋白质定量和表征的肽段的检测方法而言，合成的稳定同位素标记内标肽段至关重要。目前尚无关于在蛋白质质谱检测中使用肽段的通用建议。
美国国立癌症研究所的临床蛋白质组肿瘤分析联盟已与临床检验人员、肽段制造商、计量学家、制药行业代表及其他专业人士合作，制定了一套关于肽段采购、表征、储存和处理的共识性建议，以及对蛋白质质谱检测所产生数据的解读方法。此外，还强调了经过仔细表征的参考物质的重要性，特别是肽段标准品对于提高整个行业氨基酸分析方法一致性的重要性。围绕肽段使用的操作规范的统一以及样品制备方案的透明度应有助于实现研究和临床护理中肽段和蛋白质定量的协调一致。
