Yu Ru-Jia, Ma Wei, Liu Xiao-Yuan, Jin Hong-Ying, Han Huan-Xing, Wang Hong-Yang, Tian He, Long Yi-Tao
1. Key Laboratory for Advanced Materials, School of Chemistry & Molecular Engineering, East China University of Science and Technology, P. R. China.
2. Translational Medicine Center, Changzheng Hospital, The Second Military Medical University, P. R. China.
Theranostics. 2016 Jun 27;6(10):1732-9. doi: 10.7150/thno.16129. eCollection 2016.
Determination of disease biomarkers in clinical samples is of crucial significance for disease monitoring and public health. The dominating format is enzyme-linked immunosorbent assay (ELISA), which subtly exploits both the antigen-antibody reaction and biocatalytic property of enzymes. Although enzymes play an important role in this platform, they generally suffer from inferior stability and less tolerant of temperature, pH condition compared with general chemical product. Here, we demonstrate a metal-linked immunosorbent assay (MeLISA) based on a robust signal amplification mechanism that faithfully replaces the essential element of the enzyme. As an enzyme-free alternative to ELISA, this methodology works by the detection of α-fetoprotein (AFP), prostatic specific antigen (PSA) and C-reactive protein (CRP) at concentrations of 0.1 ng mL(-1), 0.1 ng mL(-1) and 1 ng mL(-1) respectively. It exhibits approximately two magnitudes higher sensitivity and is 4 times faster for chromogenic reaction than ELISA. The detection of AFP and PSA was further confirmed by over a hundred serum samples from hepatocellular carcinoma (HCC) and prostate cancer patients respectively.
在临床样本中测定疾病生物标志物对于疾病监测和公共卫生至关重要。主要的检测形式是酶联免疫吸附测定（ELISA），它巧妙地利用了抗原-抗体反应和酶的生物催化特性。尽管酶在这个平台中起着重要作用，但与一般化学产品相比，它们通常稳定性较差，对温度、pH条件的耐受性也较低。在此，我们展示了一种基于强大信号放大机制的金属连接免疫吸附测定（MeLISA），该机制忠实地替代了酶的关键要素。作为ELISA的无酶替代方法，该方法通过分别检测浓度为0.1 ng mL⁻¹、0.1 ng mL⁻¹和1 ng mL⁻¹的甲胎蛋白（AFP）、前列腺特异性抗原（PSA）和C反应蛋白（CRP）来发挥作用。它的灵敏度高出约两个数量级，显色反应速度比ELISA快4倍。分别来自肝细胞癌（HCC）和前列腺癌患者的一百多个血清样本进一步证实了对AFP和PSA的检测。
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