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Multianalyte immunoassay chip for detection of tumor markers by chemiluminescent and colorimetric methods

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Abstract

Most cancers developed an elevation of the level of at least two markers associated with their incidence. Simultaneous detection of multi-tumor markers associated with a particular type of cancer plays an important role in cancer diagnostic. Here, a multianalyte immunoassay chip for simple and sensitive detection of tumor markers with chemiluminescent and colorimetric methods was proposed, in which carcinoembryonic antigen (CEA) and carbohydrate antigen (CA19-9) that associated with colorectal cancer were detected as model. The immunoassay chip was fabricated by co-immobilization of CEA/CA19-9 antibody on a glass slide with γ-glycidoxypropyltrimethoxysilane as linkage. Through sandwiched immunoreactions, CEA, CA19-9, and their corresponding enzyme tracers, alkaline phosphatase-labeled anti-CEA and horseradish peroxidase-labeled anti-CA19-9, were introduced on the chip. Then, they were sequentially detected by chemiluminescent method in the range of 0.5–80 μg/L and 0.5–80 kU/L with the detection limits of 0.41 μg/L and 0.36 kU/L at 3σ for CEA and CA19-9, respectively. They could also be detected by colorimetric method in the range of 1–200 μg/L and 5–200 kU/L with the detection limits of 0.25 μg/L and 1.25 kU/L at 3σ for CEA and CA19-9, respectively. All these results demonstrated that the present work provided a promising analytical method for tumor markers’ analysis with the advantages of simple analytical procedure, small sample volume and lower cost, which made the proposed method potential for high-throughput detection.

The multianalyte immunoassay chip was fabricated for detection of tumor markers by co-immobilization of CEA/CA19-9 antibody on a glass slide with γ-glycidoxypropyltrimethoxysilane (GPMS) as linkage

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Acknowledgments

The project is supported by the National Natural Science Foundation of China (grant nos. 20875013), the Key Program (BK2010059) from the Natural Science Foundation of Jiangsu province, the Specialized Research Funds for the Doctoral Program of Higher Education (200802860035, 200802860025), Open Foundation from State Key Laboratory of Bioelectronics, and Significant Scientific Research Guidance Foundation from Southeast University.

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Authors and Affiliations

  1. School of Chemistry and Chemical Engineering, Jiangning District, Southeast University, Nanjing, 211189, China

    Wei Wei, Chunyan Zhang, Jing Qian & Songqin Liu

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  1. Wei Wei
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  2. Chunyan Zhang
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  3. Jing Qian
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  4. Songqin Liu
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Correspondence to Songqin Liu.

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Wei, W., Zhang, C., Qian, J. et al. Multianalyte immunoassay chip for detection of tumor markers by chemiluminescent and colorimetric methods. Anal Bioanal Chem 401, 3269–3274 (2011). https://doi.org/10.1007/s00216-011-5400-x

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  • DOI: https://doi.org/10.1007/s00216-011-5400-x

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