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Self-assembled protein-enzyme nanoflower-based fluorescent sensing for protein biomarker

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Abstract

Multi-protein (or enzyme) conjugates play a vital role in biosensing due to the integrated function of each component, such as biological recognition and signal amplification. In this work, a green self-assembled method for the synthesis of multi-functional protein-enzyme nanoflowers has been developed, in which no chemical modification and coupling reaction is needed to fabricate the fluorescent signal probe. The self-assembled protein-enzyme conjugates streptavidin (SA) -β-galactosidase (β-Gal)-CaHPO4 nanoflowers load sufficient enzymes without damaging their activity, which meets the requirements of signal tags for biosensing. Through integrated multi-function of biorecognition (SA) and signal amplification (β-Gal), the SA-β-Gal-CaHPO4 hybrid nanoflower-based fluorescent sensor exhibited an ultrasensitive detection of protein biomarker alpha-fetoprotein (AFP), with limits of detection at the fM level. The presented self-assembled strategy can be extensively applied to develop on-demand protein-enzyme conjugates according to the specific requirements in a variety of applications including biosensors, bioimaging, and biomedicine.

A self-assembled method has been presented for the facile and green synthesis of SA-β-Gal-CaHPO4 nanocomplexes with flower-like shape and high activity, and further employed as signal tag for fluorescent sensing of protein biomarker.

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Funding

This work was supported by the National Natural Science Foundation of China (21475101, 21675119).

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Authors and Affiliations

  1. Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, China

    Yucheng Liu, Bao Wang, Xinghu Ji & Zhike He

Authors
  1. Yucheng Liu
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  2. Bao Wang
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  3. Xinghu Ji
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  4. Zhike He
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Corresponding author

Correspondence to Zhike He.

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Ethical standards and informed consent

The study was approved by the Ethical Committee of Wuhan University. Human fluid samples used in this study do not have any identifying information about all the participants that provided written informed consent.

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The authors declare that they have no conflict of interest.

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Liu, Y., Wang, B., Ji, X. et al. Self-assembled protein-enzyme nanoflower-based fluorescent sensing for protein biomarker. Anal Bioanal Chem 410, 7591–7598 (2018). https://doi.org/10.1007/s00216-018-1398-7

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  • DOI: https://doi.org/10.1007/s00216-018-1398-7

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