Analysis of several fluorescent detector molecules for protein microarray use
Analysis of several fluorescent detector molecules for protein microarray use
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DOI：
10.1002/bio.697
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发表时间：
2003-01-01
期刊：
影响因子：
2.9
通讯作者：
Wiese, R
中科院分区：
文献类型：
--
作者：
Wiese, R
在两个蛋白质微阵列平台上评估了几种链霉亲和素连接的荧光检测分子的效用。测试的检测分子包括：Alexa Fluor 546； R-藻红蛋白 (RPE)，橙色荧光球：用 Cy3 和 RPE-抗体复合物标记的含 Cy3 的脂质体（大单层囊泡。LUV）。测试的两种阵列架构由鼠 Fc-生物素阵列和鼠 IgG 阵列组成（鼠 IgG 阵列用生物素化的兔抗鼠 IgG 进行探测）。这些平台允许通过检测器识别阵列结合的生物素来直接比较检测器的效用。所有检查的荧光检测器都在每个阵列平台上都显示出实用性。对于 Fc-生物素阵列，检测器信号强度（背景调整）如下：RPE-抗体复合物 > 荧光球 > RPE > 脂质体 > Alexa 546：对于 IgG 阵列：RPE/抗体复合物 > RPE > 荧光球 > Alexa546 > 脂质体。 RPE-抗体复合物的荧光强度分别比 Fc-生物素和鼠 IgG 阵列的下一个最接近的检测分子强 67% 和 150%。背景荧光的显着增加（与单独的 RPE 相比）并没有伴随着通过使用 RPE-抗体复合物获得的信号强度的增加（信噪比的真正增加）。这些结果表明，RPE-抗体复合物在蛋白质微阵列上分析物的荧光检测方面优于其他分子。版权所有 (C) 2002 John Wiley Sons, Ltd.
The utility of several streptavidin-linked fluorescent detector molecules was evaluated on two protein microarray platforms. Tested detector molecules included: Alexa Fluor 546; R-phycoerythrin (RPE), orange fluospheres: Cy3-containing liposomes (Large Unilamellar Vesicles. LUV) labelled with Cy3 and an RPE-antibody complex. The two array architectures tested consisted of an array of murine Fc-biotin and an array of murine IgG (the murine IgG array was probed with a biotinylated rabbit anti-murine IgG). These platforms allowed for the direct comparison of detector utility by detector recognition of array-bound biotin. All of the fluorescent detectors examined demonstrated utility on each of the array platforms. For the Fc-biotin array, detector signal intensity (background adjusted) was as follows: RPE-antibody, complex > fluospheres > RPE > liposomes > Alexa 546: for the IgG array: RPE/antibody complex > RPE > fluospheres > Alexa546 > liposomes. The RPE-antibody complex fluoresced 67% and 150% more intensely than the next closest detector molecule for the Fc-biotin and the murine IgG arrays, respectively. A marked increase in background fluorescence (as compared to RPE alone) did not accompany the increase in signal intensity gained through RPE-antibody complex use (a true increase in signal:noise ratio). These results suggest that the RPE-antibody complex is superior to other molecules for fluorescent detection of analytes on protein microarrays. Copyright (C) 2002 John Wiley Sons, Ltd.