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Entrapment of horseradish peroxidase into nanometer-scale metal–organic frameworks: a new nanocarrier for signal amplification in enzyme-linked immunosorbent assay

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Abstract

Horseradish peroxidase (HRP) was highly loaded into large holes of nanometer-scale metal–organic frameworks (i.e., PCN-333(Al)) for signal amplification in enzyme-linked immunosorbent assay (ELISA). The enzyme-labeled antibody complex prepared using nanometer-scale PCN-333(Al) maintained a high catalytic efficiency. Its Vm and Kcat values with 3,3',5,5'-Tetramethylbenzidine (TMB)-H2O2 as substrates were 4.84 × 10−5 mM/s and 4.84 × 104 min−1, respectively. We demonstrated an HRP@PCN-333 signal amplification strategy for colorimetric assay of human prostate-specific antigen (PSA). The linear range of PSA detection by using this method was 15–165 pg/mL, and the limit of detection was 6 pg/mL (S/N = 3), indicating the potential application of this method in detecting disease markers under clinical conditions. The presented strategy exhibited the characteristics of significantly increased amount of labeled enzymes, improved stability and utilization of enzymes, simple preparation process of enzyme-labeled antibodies, and low cost.

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Fig. 1
Fig. 2
Fig. 3
Fig. 4

taken from the ELISA of PSA at different standard concentrations, (B) corresponding calibration curve (■), and variation profile (□) of the detection results. (C) Calibration curve of PSA detected using an ELISA kit

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Funding

Supported by the Fundamental Research Funds for the Central Universities (GK2021001007) and the Natural Science Foundation of Shaanxi Province (2018JM2005).

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  1. Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Changan West Road 620, Xi’an, 710119, China

    Pengyue Sun, Yao Li, Jing Li & Yaodong Zhang

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  1. Pengyue Sun
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  2. Yao Li
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Pengyue Sun and Yao Li are the first authors.

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Sun, P., Li, Y., Li, J. et al. Entrapment of horseradish peroxidase into nanometer-scale metal–organic frameworks: a new nanocarrier for signal amplification in enzyme-linked immunosorbent assay. Microchim Acta 188, 409 (2021). https://doi.org/10.1007/s00604-021-05065-9

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