Multiplexed proximity ligation assays to profile putative plasma biomarkers relevant to pancreatic and ovarian cancer
Multiplexed proximity ligation assays to profile putative plasma biomarkers relevant to pancreatic and ovarian cancer
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DOI：
10.1373/clinchem.2007.093195
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发表时间：
2008-03-01
影响因子：
9.3
通讯作者：
Davis, Ronald W.
中科院分区：
文献类型：
--
作者：
Fredriksson, Simon;Horecka, Joe;Davis, Ronald W.
背景：生物标志物的发现和验证需要灵敏的方法。我们在一项分析胰腺癌和卵巢癌血浆生物标志物的初步研究中测试了一项很有前途的技术——多重接近结扎法（PLA）。方法：我们使用4组6- plex和7-plex pla检测生物标志物，每次检测消耗1 μ L血浆，使用匹配的单克隆抗体对或单批多克隆抗体。通过近距离连接将蛋白质分析物转化为独特的DNA聚合子，随后通过定量PCR检测。我们分析了18例胰腺癌病例和19例对照19例卵巢癌病例和20例对照以下蛋白质：a崩解素和金属蛋白酶8、CA-125、CA 19-9、羧肽酶A1、癌胚抗原、结缔组织生长因子、表皮生长因子受体、上皮细胞粘附分子、Her2、半乳糖凝集素-1、胰岛素样生长因子2、白细胞介素-1 α、白细胞介素-7、间皮素、巨噬细胞迁移抑制因子、骨桥蛋白、分泌性白细胞肽酶抑制剂、肿瘤坏死因子a、血管内皮生长因子、几丁质酶3-样1。CA-125探针存在于3个复刻板中。我们使用三特异性PLA，在3个探针之间进行2次结扎事件，测量了ca -125-间皮素复合物的血浆浓度。结果：检测显示CA-125的测量结果一致，与其他标记物同时检测无关，并且与Luminex数据具有良好的相关性。与文献报道相比，我们在其他假定的标记上取得了预期的结果。结论：复合PLA可采用匹配单克隆抗体或单批多克隆抗体。抗体应被证明对识别和验证假定的疾病生物标志物和发现是有用的。multimarker面板。(c) 2008美国临床化学协会。
BACKGROUND: Sensitive methods are needed for biomarker discovery and validation. We tested one promising technology, multiplex proximity ligation assay (PLA), in a pilot study profiling plasma biomarkers in pancreatic and ovarian cancer.METHODS: We used 4 panels of 6- and 7-plex PLAs to detect biomarkers, with each assay consuming 1 mu L plasma and using either matched monoclonal antibody pairs or single batches of polyclonal antibody. Protein analytes were converted to unique DNA arnplicons by proximity ligation and subsequently detected by quantitative PCR. We profiled 18 pancreatic cancer cases and 19 controls and 19 ovarian cancer cases and 20 controls for the following proteins: a disintegrin and metalloprotease 8, CA-125, CA 19-9, carboxypeptidase A1, carcinoembryonic antigen, connective tissue growth factor, epidermal growth factor receptor, epithelial cell adhesion molecule, Her2, galectin-1, insulin-like growth factor 2, interleukin-1 alpha, interleukin-7, mesothelin, macrophage migration inhibitory factor, osteopontin, secretory leukocyte peptidase inhibitor, tumor necrosis factor a, vascular endothelial growth factor, and chitinase 3-like 1. Probes for CA-125 were present in 3 of the multiplex panels. We measured plasma concentrations of the CA-125-mesothelin complex by use of a triple-specific PLA with 2 ligation events among 3 probes.RESULTS: The assays displayed consistent measurements of CA-125 independent of which other markers were simultaneously detected and showed good correlation with Luminex data. In comparison to literature reports, we achieved expected results for other putative markers.CONCLUSION: Multiplex PLA using either matched monoclonal antibodies or single batches of polyclonal. antibody should prove useful for identifying and validating sets of putative disease biomarkers and finding. multimarker panels. (c) 2008 American Association for Clinical Chemistry.