Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul 143-701, Korea.
Department of Emergency Medicine, Kyung Hee University Hospital at Gangdong, Seoul 134-727, Korea.
Sensors (Basel). 2020 Feb 24;20(4):1232. doi: 10.3390/s20041232.
A readily synthesizable fluorescent probe DMAT-π-CAP was evaluated for sensitive and selective detection of human serum albumin (HSA). DMAT-π-CAP showed selective turn-on fluorescence at 730 nm in the presence of HSA with more than 720-fold enhancement in emission intensity ([DMAT-π-CAP] = 10 μM), and rapid detection of HSA was accomplished in 3 seconds. The fluorescence intensity of DMAT-π-CAP was shown to increase in HSA concentration-dependent manner ( = 15.4 ± 3.3 μM), and the limit of detection of DMAT-π-CAP was determined to be 10.9 nM (0.72 mg/L). The 1:1 stoichiometry between DMAT-π-CAP and HSA was determined, and the displacement assay revealed that DMAT-π-CAP competes with hemin for the unique binding site, which rarely accommodates drugs and endogenous compounds. Based on the HSA-selective turn-on NIR fluorescence property as well as the unique binding site, DMAT-π-CAP was anticipated to serve as a fluorescence sensor for quantitative detection of the HSA level in biological samples with minimized background interference. Thus, urine samples were directly analyzed by DMAT-π-CAP to assess albumin levels, and the results were comparable to those obtained from immunoassay. The similar sensitivity and specificity to the immunoassay along with the simple, cost-effective, and fast detection of HSA warrants practical application of the NIR fluorescent albumin sensor, DMAT-π-CAP, in the analysis of albumin levels in various biological environments.
一种易于合成的荧光探针 DMAT-π-CAP 被评估用于灵敏和选择性检测人血清白蛋白 (HSA)。在存在 HSA 的情况下，DMAT-π-CAP 在 730nm 处显示出选择性的荧光开启，发射强度增强超过 720 倍([DMAT-π-CAP] = 10 μM)，并且可以在 3 秒内快速检测到 HSA。DMAT-π-CAP 的荧光强度显示出与 HSA 浓度依赖的增加([DMAT-π-CAP] = 15.4 ± 3.3 μM)，并且 DMAT-π-CAP 的检测限确定为 10.9 nM(0.72 mg/L)。确定了 DMAT-π-CAP 与 HSA 之间的 1:1 化学计量关系，并且置换实验表明 DMAT-π-CAP 与血红素竞争独特的结合位点，该结合位点很少容纳药物和内源性化合物。基于 HSA 选择性的近红外荧光开启特性以及独特的结合位点，预计 DMAT-π-CAP 将作为荧光传感器用于生物样品中 HSA 水平的定量检测，同时最小化背景干扰。因此，直接使用 DMAT-π-CAP 分析尿液样本以评估白蛋白水平，并且结果与免疫测定获得的结果相当。与免疫测定具有相似的灵敏度和特异性，以及对 HSA 的简单、经济高效和快速检测，证明了近红外荧光白蛋白传感器 DMAT-π-CAP 在各种生物环境中分析白蛋白水平的实际应用。
