Gold nanoparticle based immunochromatography using a resin modified micropipette tip for rapid and simple detection of human chorionic gonadotropin hormone and prostate-specific antigen
Gold nanoparticle based immunochromatography using a resin modified micropipette tip for rapid and simple detection of human chorionic gonadotropin hormone and prostate-specific antigen
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DOI：
10.1016/j.stam.2005.12.008
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发表时间：
2006-01
影响因子：
5.5
通讯作者：
Teruko Yuhi;N. Nagatani;T. Endo;K. Kerman;Masayuki Takata;H. Konaka;M. Namiki;Y. Takamura;E. Tamiya
中科院分区：
文献类型：
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作者：
Teruko Yuhi;N. Nagatani;T. Endo;K. Kerman;Masayuki Takata;H. Konaka;M. Namiki;Y. Takamura;E. Tamiya
结合硝基纤维素树脂改性的微量移液器吸头，开发了一种基于免疫层析的新型生物分析系统，即 ZipTip®。我们的生物分析系统采用了夹心式免疫测定法。将微量移液器吸出的第一抗体固定在微量移液器吸头边缘的免疫层析树脂上。封闭液也以同样的方式吸出。测定操作是通过吸出样品溶液，然后吸出金胶体纳米颗粒（Au naps）结合的二抗溶液来进行的。由于该生物分析系统使用微量移液器，因此可以增加样品体积，从而能够检测低浓度的抗原。此外，清洗程序也可以轻松进行，以降低背景水平。抗原抗体反应后，肉眼可观察到Au naps的颜色强度。对于分析评估，通过扫描仪捕获颜色强度，并通过分析软件进行处理。我们实现了人绒毛膜促性腺激素（hCG）和总前列腺特异性抗原（TPSA）的检测，这两个指标是众所周知的妊娠和癌症的基本指标。 hCG 的检测限 (LOD) 为 8 ng/ml（0.8 ng/tip），与其他基于免疫层析的传统系统相当。此外，通过应用不同的样品体积，TPSA 的 LOD 比现有系统有所改进，例如 200 μl 样品体积中的 5 ng/ml（1 ng/tip），以及 300 μl 样品体积中的 2 ng/ml（0.6 ng/tip）。由于我们的生物分析系统需要少量的固定化抗原，因此对于筛选抗原抗体反应的基础研究将非常有用。此外，我们的生物分析系统可以应用于现场筛选，因为其操作简单，并且可以快速获得直观的结果。
A novel bioanalysis system based on immunochromatography was developed in connection with a nitrocellulose resin-modified micropipette tip, namely as ZipTip®. The sandwich-type immunoassay was applied to our bioananalysis system. The first antibodies that were aspirated by the micropipette were immobilized on the immunochromatographic resin at the edge of micropipette tip. The blocking solution was also aspirated in the same fashion. The measurement operation was performed by aspirating the sample solution, and then the gold colloidal nanoparticles (Au naps) conjugated secondary antibody solution. Since this bioanalysis system utilizes a micropipette, it is possible to increase the sample volume, which would enable the detection of antigen at low concentrations. In addition, the washing procedure can also be performed easily to reduce the background level. After the antigen-antibody reaction, the color intensity of Au naps could be observed by the naked eye. For analytical evaluation, the color intensity was captured by a scanner, and processed by analysis software. We have achieved the detection of human chorionic gonadotropin hormone (hCG) and total prostate-specific antigen (TPSA), which are well-known as fundamental indicators of pregnancy and cancer. The limit of detection (LOD) for hCG was 8 ng/ml (0.8 ng/tip), which is comparable to that of other conventional systems based on immunochromatography. Moreover, the LOD for TPSA was improved over the existing systems with the application of different sample volumes, such as 5 ng/ml (1 ng/tip) in 200 μl sample volume, and 2 ng/ml (0.6 ng/tip) in 300 μl sample volume. Since our bioanalysis system requires a small amount of immobilized antigen, it would be greatly useful in basic research for screening the antigen–antibody reactions. Besides, our bioanalysis system can be applied to on-field screening, since its operation is simple, and the visual results can be obtained rapidly.