Development of a manual self-assembled colloidal gold nanoparticle-immunochromatographic strip for rapid determination of human interferon-γ
Development of a manual self-assembled colloidal gold nanoparticle-immunochromatographic strip for rapid determination of human interferon-γ
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DOI：
10.1039/c3an36547f
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发表时间：
2013-01-01
期刊：
影响因子：
4.2
通讯作者：
Chou, Shu-Fen
中科院分区：
文献类型：
--
作者：
Chou, Shu-Fen
干扰素（IFN）在抑制肿瘤生长中起作用，并参与免疫反应。在干扰素中，干扰素-γ（IFN-γ）是最重要的治疗性蛋白质之一，其免疫调节能力优于其它类型的干扰素。本研究的目的是利用抗人IFNg的多克隆抗体和单克隆抗体，开发一种手动自组装的人IFNg纳米胶体金免疫层析试纸条。由氯金酸（HAuCl 4）制成直径为25 nm的胶体金，并标记在抗IFN γ mAb上作为色度试剂。人IFNg浓度在10-1000 ng/mL范围内与像素强度呈良好的线性关系，回归方程为Y = 0.159logX + 0.0648，R-2 = 0.992; Y = 0.294logX + 0.091，R-2 = 0.9969。此外，在测定小鼠血清样品时，未发生交叉反应，检测时间约为10分钟。该试纸条在室温下的保质期在28天以上。人工操作模式的主要优点是不需要昂贵的仪器和试剂。该试纸条具有特异性强、经济、方便、不需机器等优点。
Interferons (IFNs) play a role in inhibition of tumor growth and participate in immunoreactions. Among IFNs, interferon-gamma (IFN gamma) is one of the most important therapeutic proteins and its immunodulation ability is better than that of other types. The objective of this study is to develop a manual self-assembled colloidal gold nanoparticle-immunochromatographic strip for human IFNg using anti-human IFNg polyclonal and monoclonal antibodies. Colloidal gold with a 25 nm diameter was made from chloroauric acid (HAuCl4), and labeled on anti-IFN gamma mAbs as a chrominance reagent. A good linear relationship existed between the pixel intensity and the human IFNg concentrations from 10-1000 ng mL(-1) in mouse serum and buffer, respectively, the regression equation was Y = 0.159logX + 0.0648, R-2 = 0.992 in mouse serum; Y = 0.294logX + 0.091, R-2 = 0.9969 in phosphate buffer by this proposed strip. Moreover, in the determination for mouse serum samples no cross-reaction occurred and the detection time was approximately 10 minutes. The shelf life of the strip was above 28 days at room temperature. The major advantages of the manual operation model were no expensive instruments and less reagents required. This proposed strip was highly specific, economic, convenient, and no machine was needed in clinical diagnosis.