Pancreatic cancer serum detection using a lectin/glyco-antibody array method.
Pancreatic cancer serum detection using a lectin/glyco-antibody array method.
复制标题
使用凝集素/糖抗体阵列方法检测胰腺癌血清。
DOI：
10.1021/pr8007013
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发表时间：
2009-02
影响因子：
4.4
通讯作者：
Lubman, David M.
中科院分区：
文献类型：
--
作者：
Li, Chen;Simeone, Diane M.;Brenner, Dean E.;Anderson, Michelle A.;Shedden, Kerby A.;Ruffin, Mack T.;Lubman, David M.
胰腺癌是一种可怕的疾病，需要早期发现生物标记物来改善这些患者的预后。在这项工作中，凝集素抗体微阵列被用来检测血清中蛋白质独特的糖基化模式。将先前研究中发现的四种潜在糖蛋白标志物的抗体印刷在硝酸纤维素涂层玻片上，并将这些微阵列与患者血清杂交，以提取目标糖蛋白。然后用凝集素以夹心分析的形式检测捕获的糖蛋白上不同的糖链结构单元。用来评价差异糖基化模式的生物素化凝集素有：金银花凝集素(AAL)、接骨木树皮凝集素(SNA)、山玛瑙凝集素II(MAL)、木豆凝集素(LCA)和刀豆蛋白A(ConA)。捕获的糖蛋白在微阵列上通过平板上的消化和MALDI QIT-TOF质谱学的直接分析进行原位评估。用89名正常人、35名慢性胰腺炎患者、37名糖尿病患者和22名胰腺癌患者的血清进行分析。我们发现，该方法具有良好的重复性，以对照块的信号偏差作为载玻片上的标准，并以41对纯技术重复进行测量。癌组织中α-1-β糖蛋白对凝集素SNA的反应性较其他非肿瘤组高69%(95%可信区间为53%~86%)，因此有可能以较高的灵敏度和特异性将癌症与其他疾病组和正常组织区分开来。这些数据表明，在高通量凝集素微阵列上检测到的差异糖基化模式是一种有前途的胰腺癌早期检测的生物标志物方法。
Pancreatic cancer is a formidable disease and early detection biomarkers are needed to make inroads into improving the outcomes in these patients. In this work lectin antibody microarrays were utilized to detect unique glycosylation patterns of proteins from serum. Antibodies to four potential glycoprotein markers that were found in previous studies were printed on nitrocellulose coated glass slides and these microarrays were hybridized against patient serum to extract the target glycoproteins. Lectins were then used to detect different glycan structural units on the captured glycoproteins in a sandwich assay format. The biotinylated lectins used to assess differential glycosylation patterns were Aleuria aurentia lectin (AAL), Sambucus nigra bark lectin (SNA), Maackia amurensis lectin II (MAL), Lens culinaris agglutinin (LCA), and Concanavalin A (ConA). Captured glycoproteins were evaluated on the microarray in situ by on-plate digestion and direct analysis using MALDI QIT-TOF mass spectroscopy. Analysis was performed using serum from 89 normal controls, 35 chronic pancreatitis samples, 37 diabetic samples and 22 pancreatic cancer samples. We found that this method had excellent reproducibility as measured by the signal deviation of control blocks as on-slide standard and 41 pairs of pure technical replicates. It was possible to discriminate cancer from the other disease groups and normal samples with high sensitivity and specificity where the response of Alpha-1-β glycoprotein to lectin SNA increased by 69% in the cancer sample compared to the other non-cancer groups (95% confidence interval 53% to 86%). These data suggest that differential glycosylation patterns detected on high throughput lectin microarrays are a promising biomarker approach for the early detection of pancreatic cancer.
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影响因子：
48
作者：
Chen, Songming;LaRoche, Tom;Haab, Brian B.
通讯作者：
Haab, Brian B.
影响因子：
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作者：
Drake, Richard R.;Schwegler, E. Ellen;Semmes, O. John
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Semmes, O. John
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通讯作者：
Warshaw, Andrew L.
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作者：
Ingvarsson, Johan;Wingren, Christer;Borrebaeck, Carl A. K.
通讯作者：
Borrebaeck, Carl A. K.
影响因子：
7.4
作者：
An, HJ;Peavy, TR;Lebrilla, CB
通讯作者：
Lebrilla, CB