Petersson Linn, Dexlin-Mellby Linda, Bengtsson Anders A, Sturfelt Gunnar, Borrebaeck Carl A K, Wingren Christer
Dept. of Immunotechnology and CREATE Health, Medicon Village, Lund University, Medicon Village, Building no. 406, SE-22381 Lund, Sweden.
Lab Chip. 2014 Jun 7;14(11):1931-42. doi: 10.1039/c3lc51420j. Epub 2014 Apr 24.
In the quest to decipher disease-associated biomarkers, miniaturized and multiplexed antibody arrays may play a central role in generating protein expression profiles, or protein maps, of crude serum samples. In this conceptual study, we explored a novel, 4-times larger pen design, enabling us to, in a unique manner, simultaneously print 48 different reagents (antibodies) as individual 78.5 μm(2) (10 μm in diameter) sized spots at a density of 38,000 spots cm(-2) using dip-pen nanolithography technology. The antibody array set-up was interfaced with a high-resolution fluorescent-based scanner for sensitive sensing. The performance and applicability of this novel 48-plex recombinant antibody array platform design was demonstrated in a first clinical application targeting SLE nephritis, a severe chronic autoimmune connective tissue disorder, as the model disease. To this end, crude, directly biotinylated serum samples were targeted. The results showed that the miniaturized and multiplexed array platform displayed adequate performance, and that SLE-associated serum biomarker panels reflecting the disease process could be deciphered, outlining the use of miniaturized antibody arrays for disease proteomics and biomarker discovery.
在探寻与疾病相关的生物标志物的过程中，小型化和多重抗体阵列可能在生成粗血清样本的蛋白质表达谱或蛋白质图谱方面发挥核心作用。在这项概念性研究中，我们探索了一种新颖的、尺寸大四倍的笔式设计，这使我们能够以独特的方式，使用蘸笔纳米光刻技术，以每平方厘米38,000个点的密度，将48种不同试剂（抗体）同时打印成直径为10μm、面积为78.5μm²的单个斑点。抗体阵列装置与基于高分辨率荧光的扫描仪相连以进行灵敏检测。这种新型的48重重组抗体阵列平台设计的性能和适用性在首个以狼疮性肾炎（一种严重的慢性自身免疫性结缔组织疾病）为模型疾病的临床应用中得到了验证。为此，以未经处理的、直接生物素化的血清样本为检测对象。结果表明，小型化和多重阵列平台表现出了足够的性能，并且能够解读反映疾病进程的与系统性红斑狼疮相关的血清生物标志物组，勾勒出了小型化抗体阵列在疾病蛋白质组学和生物标志物发现中的应用。
