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Development of indirect competitive immunoassay for highly sensitive determination of ractopamine in pork liver samples based on surface plasmon resonance sensor

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文献类型：外文期刊

作者： Liu, Ming ¹ ; Ning, Baoan ¹ ; Qu, Lijie ² ; Peng, Yuan ¹ ; Dong, Jianwei ² ; Gao, Na ² ; Liu, Lei ³ ; Gao, Zhixian ¹ ;

作者机构： 1.Inst Hlth & Environm Med, Tianjin Key Lab Risk Assessment & Control Technol, Tianjin 300050, Peoples R China

2.Huazhong Agr Univ, Coll Food Sci & Technol, Wuhan 430070, Peoples R China

3.Tianjin Acad Agr Sci, Cent Lab, Tianjin 300381, Peoples R China

关键词： Condition optimization;Immunosensor;Indirect competitive assay;Ractopamine;Surface plasmon resonance

期刊名称：SENSORS AND ACTUATORS B-CHEMICAL （影响因子：7.46；五年影响因子：6.743 ）

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收录情况： SCI

摘要： In this work, we demonstrate the development of a highly sensitive and selective surface plasmon resonance (SPR) immunosensor for label-free detection of ractopamine (RCT) as low as 0.12 ppb (ng mL ~(-1)) direct from pork liver samples. RCT is a kind of the β-adrenergic agonist with N-alkylphenyl substituent, and the development of high performance sensing methods for RCT is a critical issue because of its vital role in human metabolism. To construct the immunosensor system, the biosensor surface was formed by immobilization of ractopamine-ovalbumin (RCT-OVA) conjugate onto an Au-thiolate self-assembled SPR sensor chip by covalent amide binding, and the detection of RCT was performed based on indirect competitive inhibition principle. Detailed works were carried out focusing on optimizing the conditions that affect immunosensitivity, including immobilization buffer, immunobinding buffer, RCT-OVA concentration, antibody concentration, activation time, regeneration buffer, etc. A preferable way to determine the optimal antibody concentration was performed comparing with that used in most other reported researches. Under the optimal conditions, the biosensor exhibits a linear detection range of 0.28-4.29 ng mL ~(-1), LOD of 0.12 ng mL ~(-1) and IC 50 of 1.17 ng mL ~(-1). It improved that the developed SPR immunosensor is more sensitive for detection of RCT against other reported ones.

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