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Determination of alkaline phosphatase activity and of carcinoembryonic antigen by using a multicolor liquid crystal biosensor based on the controlled growth of silver nanoparticles

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Abstract

An ultrasensitive liquid crystal biosensor is described for multicolor visualization of the activity of alkaline phosphatase (ALP) based on the controlled growth of silver nanoparticles. The enzymatic product is accumulated on the surface of the LC sensing film by means of silver deposition, and the birefringent signal (observed with a polarizing microscope) is strongly enhanced as a result. The presence of AuNPs also enhances the sensitivity by about 4 orders of magnitude. The bright spots in polarized optical microscopy (POM) images increase with increasing activities of ALP. The signal intensities of the spots are then calculated by using Photoshop software and by multiplying the average brightness of the spots by the pixel value. The detection limit for ALP is 1.2 nU·mL−1, which is 5–7 orders of magnitude lower than other colorimetric or fluorometric methods. The method was applied to a highly sensitive immunoassay for the carcinoembryonic antigen (CEA) by integrating immunomagnetic separation. The immunoassay was applied to the analysis of complex samples without tedious sample pretreatment, and a detection limit as low as 0.35 pg·mL−1 of CEA was achieved. The method has attractive features in that it provides an ultrasensitive multicolor visualization approach for enzymes such as ALP, but also paves the way to a new kind of immunoassay coupled to immunomagnetic separation.

A signal enhanced liquid crystal (LC)-based multicolor immunosensor is described that is based on immunomagnetic separation and biometallization. Alkaline phosphatase (ALP) and carcinoembryonic antigen (CEA) can be easily visualized by bare eyes using the polarized optical microscopy (POM) images of LCs.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (NSFC, 21505119, 21465025), Applied and Basic Research Program of Yunnan Province (NO. 2015FD002), the Yunnan University’s Research Innovation Fund for Graduate Student (NO. 2018Z068). The authors would also like to thank Prof. Xiao Hong Chen at Yunnan University for her kindly support on this research.

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  1. Chuan-Hua Zhou and Qin-jiang Zi contributed equally to this work.

Authors and Affiliations

  1. School of Chemical Science and Technology, Functional Molecules Analysis and Biotransformation Key laboratory of Universities in Yunnan Province, Key Laboratory of Medicinal Chemistry for Natural Resource (Ministry of Education), Yunnan University, Kunming, 650091, China

    Chuan-Hua Zhou, Qin-jiang Zi, Jin Wang, Wen-Ying Zhao & Qiue Cao

  2. Eryuan Institute for Food and Drug Control, Dali, 671200, China

    Qin-jiang Zi

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  1. Chuan-Hua Zhou
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  2. Qin-jiang Zi
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  3. Jin Wang
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  4. Wen-Ying Zhao
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  5. Qiue Cao
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Correspondence to Chuan-Hua Zhou or Qiue Cao.

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Zhou, CH., Zi, Qj., Wang, J. et al. Determination of alkaline phosphatase activity and of carcinoembryonic antigen by using a multicolor liquid crystal biosensor based on the controlled growth of silver nanoparticles. Microchim Acta 186, 25 (2019). https://doi.org/10.1007/s00604-018-3131-9

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