Citation: ZHANG Lan, LI Xiao-Qin, WANG Guang-Li. Photoswitching Enzymatic Activity of Horseradish Peroxidase by Graphite Phase Carbonitride for Colorimetric Immunoassay[J]. Chinese Journal of Analytical Chemistry, ;2020, 48(6): 703-711. doi: 10.19756/j.issn.0253-3820.201065 shu

Photoswitching Enzymatic Activity of Horseradish Peroxidase by Graphite Phase Carbonitride for Colorimetric Immunoassay

Key Laboratory of Synthetic and Biological Colloids, Ministry of Education, School of Chemical and Material Engineering, Jiangnan University, Wuxi 214122, China

Received Date: 10 February 2020
Revised Date: 23 March 2020

Fund Project: This work was supported by the National Natural Science Foundation of China (Nos. 21575052, 21676123).

The g-C₃N₄ was found to active the activity of horseradish peroxidase (HRP) for catalyzing the oxidation of the typical substrate of 3,3',5,5'-tetramethylbenzidine (TMB) under visible light irradiation. Based on this, a novel sandwich-type immunosensor colorimetric strategy was developed for the highly sensitive detection of α-fetoprotein (AFP). In this assay, the capture antibodies (Ab₁) attached to the 96-well plates was used to capture the target of AFP, and then reacted with the detection antibody (Ab₂) and HRP co-immobilized gold nanoparticles (AuNPs) (i.e., the bioconjugates of the Ab₂-AuNPs-HRP), which was assisted by the formation of the tyramide-HRP repeats to amplify the signal. The addition of the g-C₃N₄ activated the HRP under visible light for oxidizing TMB, producing the absorption signal at 652 nm (for oxidized TMB) that correlated to the concentrations of AFP in the range from 0.5 pg/mL to 0.1 μg/mL. A limit of detection (LOD) of 0.17 pg/mL was obtained (S/N=3). The developed immunoassay could be facilely operated with wide detection range and high sensitivity, showing great promise for real sample detection.
- Graphite-phase carbon nitride,
- Horseradish peroxidase,
- Activation,
- Immunoassay
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