Detection of alpha-fetoprotein through biological signal amplification by biosensor based on imaging ellipsometry |
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Authors: | Chenghong Huang Yanyan Chen Chunxia WangWei Zhu Hongwei MaGang Jin |
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Affiliation: | a Suzhou Institute of Nano-tech and Nano-bionics, Chinese Academy of Sciences, Suzhou, Chinab NML, Institute of Mechanics, Chinese Academy of Sciences, Beijing, Chinac Institute of Biophysics, Chinese Academy of Sciences, Beijing, Chinad Graduate School of the Chinese Academy of Sciences, Beijing, Chinae Radiation Medical Institute of Shandong Academy of Medical science, Jinan, China |
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Abstract: | High sensitive and quantitative detection of alpha-fetoprotein (AFP) by biosensor based on imaging ellipsometry (BIE) through biological amplification was investigated. AFP firstly reacted with the rat monoclonal antibody (rat-mAb) initially immobilized on glutaraldehyde modified silicon surface, then rabbit anti-human AFP polyclonal antibodies (Rabbit-pAb) and goat anti-rabbit IgG (goat-IgG) were sequentially applied to amplify signal. Results revealed that signal was enhanced approximately six fold. The linear range of AFP detection was 20.0-200.0 ng/ml with a low limit of 5.0 ng/ml (S/N = 3). The cross-reaction rate was less than 5.2% evaluated by biomarker (carcinoembryonic antigen, carbohydrate 19-9 and carbohydrate antigen 242) and two common proteins (human serum albumin, fibrinogen) and their mixture. Coefficient variation (CV) for intra-slide and inter-slide reproducibility were 10.3%, 6.6%, 6.3% and 10.7%, 7.9%, 6.4% for 41.6 ng/ml, 83.2 ng/ml and 128.4 ng/ml AFP. In clinic application, cut-off value at 28.6 ng/ml was also determined with sensitivity 0.72 and specificity 0.94. Results of 47 clinic patient samples detected by BIE were in good agreement with those of electrochemiluminescence immunoassay (ECLIA) (R2 = 0.9949).The area under receiver operating characteristic curve (ROC) curve is 0.89. Statistical analysis showed that BIE is high agreement with ECLIA (Kappa = 0.733, U = 3.21>U0.01). It shows a potential for hepatocellular carcinoma (HCC) diagnosis. |
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Keywords: | Imaging ellipsometry Biosensor Signal amplification Alpha-fetoprotein Hepatocellular carcinoma |
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