Fabrication of an Electrochemical Sensor for Phospholipase C Detection Based on Flexible Electrodes and Signal Amplification Technology

doi:10.19894/j.issn.1000-0518.240026

Chinese Journal of Applied Chemistry ›› 2024, Vol. 41 ›› Issue (7): 987-997.DOI: 10.19894/j.issn.1000-0518.240026

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Fabrication of an Electrochemical Sensor for Phospholipase C Detection Based on Flexible Electrodes and Signal Amplification Technology

Yao-Yao WANG, Ming-Yang SUN, Zuan YANG, Shao-Kai DU, Yu-Xuan HE, Yue SUN()

College of Chemistry and Chemical Engineering，Liaoning Normal University，Dalian 116029，China

Received:2024-01-23 Accepted:2024-05-21 Published:2024-07-01 Online:2024-08-03
Contact: Yue SUN
About author:yue0411@163.com
Supported by:
the Natural Science Foundation of Liaoning Province(2021-MS-273);the Service Local Projects of Liaoning Education Department(JYTMS20231091);the National College Students Innovation and Entrepreneurship Training Program(202210165027);the Innovation and Entrepreneurship Training Program for College Students in Liaoning Province(S202310165008X)

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Abstract

Abstract:

Phospholipase C （PLC） is an important lipid hydrolase in plants and animals， which plays an important role in the metabolism， growth， inflammation， differentiation， senescence， and apoptosis of mammalian cells， and is significantly associated with the onset and progression of many cancers. Protein-catalyzed atom transfer radical polymerization （PATRP） is an effective method of designing and preparing macromolecules， and it can be used to link hundreds of monomers by a chain reaction to form a high-molecular-mass polymer， so that the molecular recognition signals are amplified hundreds of times. In this experiment， gold nanoparticles （Au NPs） were first deposited on the surface of flexible carbon cloth （CC） electrode， and then the PLC-specific substrate phosphatidylethanolamine （PE） was immobilized to the electrode by 3-mercaptopropionic acid， carbodiimide hydrochloride and n-hydroxysuccinimide. After the phosphoric acid group was produced by the specific hydrolysis of PE by PLC， and Zr⁴⁺ was introduced， the polymerization initiator α-bromophenylacetic acid was modified to the electrode surface by the coordination of Zr⁴⁺. Finally， PATRP of 2，2，6，6-tetramethyl-1-piperidinyloxy （TEMPO） was carried out， and electroactive polymers chains were introduced to the electrode surface. Thus a signal amplification electrochemical sensor ［P（TEMPO）/PE/Au NPs/CC］ was fabricated to detect PLC with high sensitivity. The results show that the sensor detected PLC with a linear range of 10~1×10⁵ mU/L and a detection limit of 3.568 mU/L （S/N=3）. Compared with other methods for the detection of PLC， the sensor has a wider detection range and lower detection limit， and it has a huge practical application potential in detecting PLC in real breast cancer cells.

Key words: Phospholipase C, Atom transfer radical polymerization, Flexible electrode, Electrochemical sensors

CLC Number:

O655

Yao-Yao WANG, Ming-Yang SUN, Zuan YANG, Shao-Kai DU, Yu-Xuan HE, Yue SUN. Fabrication of an Electrochemical Sensor for Phospholipase C Detection Based on Flexible Electrodes and Signal Amplification Technology[J]. Chinese Journal of Applied Chemistry, 2024, 41(7): 987-997.

Figures/Tables 10

Fig.1 Fabrication scheme of P（TEMPO）/PE/AuNPs/CC electrode via PATRP

Fig.2 SEM of bare carbon cloth （CC） electrode （A）， AuNPs/CC electrode （B）， PE/AuNPs/CC electrode （C） and P（TEMPO）/PE/AuNPs/CC electrode （D）

Fig.3 XPS spectra of P（TEMPO）/PE/AuNPs/CC electrode （A） and high-resolution spectra of Au （B）， Zr （C） and Br （D）

Fig.4 The CV of different modified electrodes in 0.1 mol/L KCl+5 mmol/L ［Fe（CN）6］3-/4- solution

Fig.5 （A） SWV curves of sensor in 0.5 mol/L LiClO4 solution to detect PLC with different concentrations （the curves from a to f correspond to PLC concentrations of 10， 1×102， 1×103， 1×104， 1×104.69 and 1×105 mU/L）；（B） Working curve of PLC detected by sensor

Table 1 Performance comparison with other analysis methods

Detection method	Main materials	Linear range/ （nmol·L^-1）	Detection limit/ （nmol·L^-1）	Ref.
Fluorescence method	11-Mercaptocarboxylic acid-gold nanodot-liposome hybrids	0.5~50	0.21	［43］
Fluorescence method	Fluorescently labeled liposomes	6.75~4.05×10²	2.7	［44］
Liquid mass spectrometry	1-Palmitoyl-2-oleoyl-diacylglycerol	0~27.5	0.4	［45］
Fluorescence method	Water-soluble conjugated polyelectrolyte-lipid complexes	0~1×10⁵	1	［46］
Electrochemical method	P（TEMPO）/PE/AuNPs/CC-modified electrodes	1.35×10^-2~1.35×10²	4.817×10^-3	This work

Fig.6 Reproducibility of P（TEMPO）/PE/AuNPs/CC electrodes

Fig.7 Selectivity of the sensor to different substances

Fig.8 Stability of the sensor

Fig.9 Effect of temperature on sensor stability

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Fabrication of an Electrochemical Sensor for Phospholipase C Detection Based on Flexible Electrodes and Signal Amplification Technology

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