NanoProbeArrays for the Analysis of Ultra-Low-Volume Protein Samples Using Piezoelectric Liquid Dispensing Technology
NanoProbeArrays for the Analysis of Ultra-Low-Volume Protein Samples Using Piezoelectric Liquid Dispensing Technology
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DOI：
10.1016/j.jala.2010.07.005
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发表时间：
2011-04-01
期刊：
影响因子：
--
通讯作者：
Wiktor, Peter
中科院分区：
文献类型：
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作者：
Nagaraj, Vinay Janthakahalli;Eaton, Seron;Wiktor, Peter
抗体微阵列作为一种高通量的蛋白质组研究技术正日益受到人们的欢迎。然而，从大多数体液或活检样本中提取的蛋白质体积非常小，通常不适合大规模抗体微阵列研究。为了证明仅用几纳升样品进行蛋白质分析的潜力，我们开发了一种名为NanoProbeArrays的新技术，该技术基于压电液体分配，用于非接触式打印和探测抗体阵列。在传统的微阵列筛选中，蛋白质样品被淹没在抗体微阵列表面，而压电喷墨打印机被用来在阵列上的抗体点上分配纳米升荧光标记的蛋白质。NanoProbeArrays能够精确地识别和可靠地区分来自不同来源的测试蛋白，而与传统的抗体微阵列相比，没有任何灵敏度和特异性的损失。通过检测血清中的细胞因子白介素-4，测试了NanoProbeArrays在复杂生物样品中生物标志物鉴定的实用性。与传统抗体微阵列相比，NanoProbeArray分析过程中样品体积的显著减少，为基础和应用蛋白质组学研究提供了新的机会。(比特杰拉2011;16:126-33)
Antibody microarrays are gaining popularity as a high-throughput technology to investigate the proteome. However, protein extracts from most body fluid or biopsy samples are available in very small volumes and are often unsuitable for large-scale antibody microarray studies. To demonstrate the potential for protein analysis with as little as a few nanoliters of sample, we have developed a new technology called NanoProbeArrays based on piezoelectric liquid dispensing for non-contact printing and probing of antibody arrays. Instead of flooding the protein sample on the antibody microarray surface, as in conventional microarray screening, a piezoelectric inkjet printer is used to dispense nanoliters of fluorescently labeled proteins over the antibody spots on the array. The ability of NanoProbeArrays to precisely identify and reliably distinguish between test proteins from different sources, without any loss of sensitivity and specificity as compared with conventional antibody microarrays, is illustrated here. The utility of NanoProbeArrays for biomarker identification in a complex biological sample was tested by detecting the cytokine interleukin-4 in serum. The significant reduction in volume of sample during NanoProbeArray analysis, as compared with conventional antibody microarrays, offers new opportunities for basic and applied proteomic research. (JALA 2011;16:126-33)