Sandwich-type, antibody microarrays for the detection and quantification of cardiovascular risk markers
Sandwich-type, antibody microarrays for the detection and quantification of cardiovascular risk markers
复制标题
DOI：
10.1016/j.snb.2007.03.005
复制
发表时间：
2007-08-08
影响因子：
8.4
通讯作者：
Gurbuz, Yasar
中科院分区：
文献类型：
--
作者：
Gul, Ozgur;Calay, Ediz;Gurbuz, Yasar
研制了夹心型抗体芯片Tay，用于包括C反应蛋白在内的心血管危险标志物的检测和定量。肿瘤坏死因子-α、血清淀粉样蛋白A。脂肪酸结合蛋白。在这项试验中，心血管危险标记蛋白抗原的抗体被固定在超环氧涂层玻璃辅助材料上作为微阵列。然后用纯抗原或人血清探测这些微阵列，然后用生物素标记的检测抗体进行探测。与荧光链霉亲和素反应后，扫描生物素标记的抗体并进行荧光定量。结果表明，C-反应蛋白、肌红蛋白、肿瘤坏死因子-α、血清淀粉样蛋白-A的基础水平和升高水平可以被检测和量化。这项研究是此类研究的第一次，利用夹心型阵列检测和量化人类血清中的多个心血管危险标志物。在这项研究中，我们还发现，作为捕获分子的抗脂肪酸结合蛋白抗体与微阵列平台上的所有检测抗体发生交叉反应，产生噪声信号，干扰标志物的检测和定量。最后，我们将我们的研究结果与市售的酶联免疫吸附试验试剂盒的性能进行了比较，这些试剂盒具有能够同时检测和定量多个标记物的优点，并且比一些商用的酶联免疫吸附试验试剂盒的动态范围更好。(C)2007 Elsevier B.V.保留所有权利。
A sandwich-type antibody microarray Tay was developed for the detection and quantitation of cardiovascular risk markers including C-reactive protein. TNF-alpha, serum amyloid-A. fatty acid binding protein. In this assay, antibodies to cardiovascular risk marker protein antigens were immobilized on super-epoxy coated glass Aides as microarrays. These microarrays were then probed with pure antigens or human sera followed by biotinylated detection antibodies. Biolinylated antibodies were scanned and quantitated for flourescence after reaction with flourescent streptavidin. Results indicate that baseline and elevated levels of C-reactive protein, myoglobin, TNF-alpha serum amyloid-A can be detected and quantified. This study is the first study of its kind, utilizing sandwich-type arrays for the detection and quantification of multiple-cardiovascular risk markers in human serum. We have also presented, in this study, that anti-fatty-acid binding protein antibodies, used as capture molecules, cross reacts with all detection antibodies on the microarray platform, generating noise signals, interfering the detection and quantification of markers. Finally, we have compared the results of our study with the performance of commercially available ELISA test-kits that presenting the advantages such as being able to detect and quantify multiple markers at once and also better dynamic ranges over some of the commercial ELISA test-kits. (c) 2007 Elsevier B.V. All rights reserved.