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Ultrasensitive multiplex SERS immunoassay based on porous Au–Ag alloy nanoparticle–amplified Raman signal probe and encoded photonic crystal beads

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Abstract

An ultrasensitive multiplex surface-enhanced Raman scattering (SERS) immunoassay was developed using porous Au–Ag alloy nanoparticles (p-AuAg NPs) as Raman signal amplification probe coupling with encoded photonic crystal microsphere. p-AuAg NPs were synthesized and modified with the second antibody (Ab2) and Raman tag (mercaptobenzoic acid, MBA) to prepare a Raman signal–amplified probe. The high porosity of the p-AuAg NPs enables significant coupling of the localized surface plasmon resonance and thus abundant inherent hotspots for Raman signal enhancement. 3D-ordered silver nanoparticles–coated silica photonic crystal beads (Ag/SPCBs) were prepared as encoded SERS substrate for multiplex detection using their reflection peaks. The signal-amplified probe was used for multiplex detection of tumor markers carcinoembryonic antigen (CEA) and alpha fetoprotein (AFP). The wide linear ranges of 10−7–103 ng/mL for CEA and 10−4–103 ng/mL for AFP with detection limits of 1.22 × 10−8 ng/mL and 2.47 × 10−5 ng/mL for CEA and AFP at a signal-to-noise ratio of 3 were obtained. The proposed multiplex SERS immunoassay method displays ultrahigh sensitivity, wide linear range, and excellent specificity, which can be successfully applied to measure clinical serum samples with satisfactory results. The research provides a novel SERS signal enhancement strategy for the multiplex bioassay.

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Funding

The authors thank the financial support from the National Natural Science Foundation of China (21575125, 21475116), the Natural Science Foundation of Jiangsu Province (BK20221370, BK20191434), Project for Science and Technology of Yangzhou (YZ2022074, YZ2020067), Key University Natural Science Foundation of Jiangsu Province (20KJA150004), Zhejiang Provincial Natural Science Foundation of China (LY20B050008), and Priority Academic Program Development of Jiangsu Higher Education Institution (PAPD).

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  1. Huizhen Yang, Jiayin Li, and Yan Rao contributed to this work equally.

Authors and Affiliations

  1. College of Chemistry and Chemical Engineering, Yangzhou University, Yangzhou, 225002, People’s Republic of China

    Huizhen Yang, Jiayin Li, Yan Rao, Linan Yang, Yu Zhang, Zhanjun Yang & Juan Li

  2. Jinhua Hospital, Zhejiang University School of Medicine, Hangzhou, China

    Yadong Xue

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  1. Huizhen Yang
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Yang, H., Li, J., Rao, Y. et al. Ultrasensitive multiplex SERS immunoassay based on porous Au–Ag alloy nanoparticle–amplified Raman signal probe and encoded photonic crystal beads. Microchim Acta 190, 13 (2023). https://doi.org/10.1007/s00604-022-05539-4

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