Citation: HE Feng, WANG Hai-Jie, LI Teng-Fei, DU Peng-Fei, WANG Wei-Ting, WANG Shou-Jing, LIU Yao-Bo, MA Yan-Li, HU Peng, TAN Tian-Yu, HAN Qing. Magnetic Separation Immunosensor Based on Enzyme-linked Gold Nanocomposite Probe for Detection of Ractopamine[J]. Chinese Journal of Analytical Chemistry, ;2023, 51(1): 102-111. doi: 10.19756/j.issn.0253-3820.221241 shu

Magnetic Separation Immunosensor Based on Enzyme-linked Gold Nanocomposite Probe for Detection of Ractopamine

HE Feng ^1,2 ,
WANG Hai-Jie ^2,3 ,
LI Teng-Fei ^1,, ,
DU Peng-Fei ^2,, ,
WANG Wei-Ting ² ,
WANG Shou-Jing ² ,
LIU Yao-Bo ² ,
MA Yan-Li ² ,
HU Peng ² ,
TAN Tian-Yu ¹ ,
HAN Qing ^1,2

1.
School of Life Sciences and Food Engineering, Hebei University of Engineering, Handan 056038, China;
2.
Key Laboratory of Novel Food Resources Processing, Ministry of Agriculture, Shandong Provincial Key Laboratory of Agro-Products Processing Technology, Institute of Agro-Food Sciences and Technology, Shandong Academy of Agricultural Sciences, Jinan 250100, China;
3.
College of Life Sciences, Yantai University, Yantai 264005, China

Corresponding author: LI Teng-Fei, DU Peng-Fei,
Received Date: 16 May 2022
Revised Date: 7 October 2022

Fund Project: Supported by the Natural Science Foundation of Shandong Province, China (No.ZR2020QC250), the Agriculture Industry Technology System (No.CARS-38), the Modern Agricultural Technology Industry System of Shandong Province, China (No.SDAIT-10-10) and the Key Research and Development Projects of Hebei Province, China (No.22325501D).

A magnetic separation competitive immunosensor based on novel enzyme-linked gold nanocomposite probes (E-GNPs) was constructed for detection of ractopamine (RAC). Through the specific interaction between streptavidin and biotin, horseradish peroxidase (HRP) was labeled on the antibody that was modified on the surface of gold nanoparticles (AuNPs) by electrostatic assembly method to obtain E-GNPs. Ovalbumin (OVA)-RAC hapten coated magnetic beads (MBs) was introduced for a competitive reaction, and quantitative analysis of the target was achieved by a color reaction. With the change of the concentration of target molecule RAC, the color of the enzyme-catalyzed substrate changed, and the concentration of RAC had a linear relationship with the color signal intensity. The UV-vis absorption spectroscopy characterization results showed that one E-GNP could carry 11 HRP molecules, which made the immunosensor highly sensitive. The prepared sensor was successfully used for detection of RAC with a detection limit of 1.75 pg/mL, showing 10 times higher sensitivity than that of traditional ELISA method. Cross-reaction experiments showed that the sensor had good selectivity for RAC, and could be applied to real samples such as pork, beef and mutton, with spiked recoveries ranging from 88.3% to 103.4%. This analytical method provided a new idea for rapid screening of RAC in animal-derived foods.
- Ractopamine,
- Gold nanoparticles,
- Magnetic microbeads,
- Horseradish peroxidase
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