Multiplexed quantification of estrogen receptor and HER2/Neu in tissue and cell lysates by peptide immunoaffinity enrichment mass spectrometry.
Multiplexed quantification of estrogen receptor and HER2/Neu in tissue and cell lysates by peptide immunoaffinity enrichment mass spectrometry.
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DOI：
10.1002/pmic.201100587
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发表时间：
2012-04
期刊：
影响因子：
3.4
通讯作者：
Paulovich, Amanda G.
中科院分区：
文献类型：
--
作者：
Schoenherr, Regine M.;Whiteaker, Jeffrey R.;Zhao, Lei;Ivey, Richard G.;Trute, Mary;Kennedy, Jacob;Voytovich, Uliana J.;Yan, Ping;Lin, Chenwei;Paulovich, Amanda G.
获得更广泛的定量蛋白质分析将对指导疾病治疗的组织标记物的数量和使用产生重大影响。近年来，基于定量质谱学的多肽和蛋白质分析(如免疫SRM分析)在血浆或尿液等生物体液中蛋白质定量方面的应用有了巨大的增长。在这里，我们通过展示多重免疫-SRM方法的应用来扩展该技术的能力，以定量测定细胞系裂解物和人类手术标本中雌激素受体(ER)和人表皮生长因子受体2(HER2)的水平。用多肽响应曲线来表征分析的性能，线性范围覆盖大约4个数量级，检测限在低fmol/mg裂解物范围内。重复性可以接受，中位变异系数约为10%。我们将该方法应用于基于ER/HER2状态的特征良好的细胞系裂解物中ER和HER2的测量。最后，对手术切除的乳腺癌组织中的蛋白质进行了检测，结果显示与临床检测确定的ER/HER2状态有很好的相关性。这是首次在细胞裂解物和人类手术标本中实施基于多肽的免疫SRM检测技术。
Access to a wider range of quantitative protein assays would significantly impact the number and use of tissue markers in guiding disease treatment. Quantitative mass spectrometry-based peptide and protein assays, such as immuno-SRM assays, have seen tremendous growth in recent years in application to protein quantification in biological fluids such as plasma or urine. Here, we extend the capability of the technique by demonstrating the application of a multiplexed immuno-SRM assay for quantification of estrogen receptor (ER) and human epidermal growth factor receptor 2 (HER2) levels in cell line lysates and human surgical specimens. The performance of the assay was characterized using peptide response curves, with linear ranges covering approximately 4 orders of magnitude and limits of detection in the low fmol/mg lysate range. Reproducibility was acceptable with median coefficients of variation of approximately 10%. We applied the assay to measurements of ER and HER2 in well-characterized cell line lysates with good discernment based on ER/HER2 status. Finally, the proteins were measured in surgically resected breast cancers, and the results showed good correlation with ER/HER2 status determined by clinical assays. This is the first implementation of the peptide-based immuno-SRM assay technology in cell lysates and human surgical specimens.
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影响因子：
158.5
作者：
Lynch, TJ;Bell, DW;Haber, DA
通讯作者：
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通讯作者：
Carr, Steven A.