Detection of protein biomarkers using RNA aptamer microarrays and enzymatically amplified surface plasmon resonance imaging
Detection of protein biomarkers using RNA aptamer microarrays and enzymatically amplified surface plasmon resonance imaging
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DOI：
10.1021/ac061849m
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发表时间：
2007-02-01
影响因子：
7.4
通讯作者：
Corn, Robert M.
中科院分区：
文献类型：
--
作者：
Li, Yuan;Lee, Hye Jin;Corn, Robert M.
开发了一种用于检测皮摩尔浓度的蛋白质生物标志物的方法，该方法利用RNA适体微阵列的表面等离子体共振成像（SPRI）测量。通过形成表面适体-蛋白质-抗体复合物来检测蛋白质在RNA微阵列上的吸附。然后使用由与抗体缀合的辣根过氧化物酶催化的局部沉淀反应来放大SPRI响应信号。这种酶促扩增SPRI方法首先通过检测浓度为500 fM的人凝血酶来表征;从三种潜在凝血酶适体候选物的微阵列中鉴定用于夹心测定的适当凝血酶适体。然后使用SPRI方法检测1 pM生物相关浓度的蛋白质血管内皮生长因子（VEGF）。VEGF是一种信号蛋白，已被用作类风湿性关节炎、乳腺癌、肺癌和结肠直肠癌的血清生物标志物，并且还与年龄相关性黄斑变性相关。
A methodology for the detection of protein biomarkers at picomolar concentrations that utilizes surface plasmon resonance imaging (SPRI) measurements of RNA aptamer microarrays is developed. The adsorption of proteins onto the RNA microarray is detected by the formation of a surface aptamer-protein-antibody complex. The SPRI response signal is then amplified using a localized precipitation reaction catalyzed by the enzyme horseradish peroxidase that is conjugated to the antibody. This enzymatically amplified SPRI methodology is first characterized by the detection of human thrombin at a concentration of 500 fM; the appropriate thrombin aptamer for the sandwich assay is identified from a microarray of three potential thrombin aptamer candidates. The SPRI method is then used to detect the protein vascular endothelial growth factor (VEGF) at a biologically relevant concentration of 1 pM. VEGF is a signaling protein that has been used as a serum biomarker for rheumatoid arthritis, breast cancer, lung cancer, and colorectal cancer and is also associated with age-related macular degeneration.