Xia Wanqiu, Zhang Lei, Wang Jianping
College of Veterinary Medicine, Hebei Agricultural University, Baoding 071000, China.
Veterinary Biological Technology Innovation Center of Hebei Province, Baoding 071000, China.
Foods. 2023 Jan 7;12(2):284. doi: 10.3390/foods12020284.
In this study, the genes of ribosomal protein S12 and renilla luciferase were linked and expressed to produce a fusion protein, and its intermolecular interactions and affinities with sevenaminoglycosides were studied. Then, the fusion protein was used as the core agent to develop a bioluminescent method on a conventional microplate for determination of the residues of thesevenaminoglycosides in pork. This method contained only one sample-loading step, and thus the assay was finished within 30 min. The limits of detection for the sevendrugs were in the range of 0.51-1.1 ng/mL, and the sensitivity for a specific drug was mainly determined by the receptordrug affinity but not related with the binding energy. After general comparison, the present method showed generally better performances than the previously reported enzyme-linked immunosorbent assays for aminoglycosides. This is the first study reporting the recognition mechanisms of ribosomal protein S12 for aminoglycosides and developing a bioluminescent method for detection of aminoglycoside residues in pork samples.
在本研究中，核糖体蛋白S12基因与海肾荧光素酶基因相连并表达以产生融合蛋白，研究了其分子间相互作用以及与七种氨基糖苷类药物的亲和力。然后，以该融合蛋白为核心试剂，在传统微孔板上建立了一种生物发光法，用于测定猪肉中七种氨基糖苷类药物的残留量。该方法仅包含一个加样步骤，因此检测可在30分钟内完成。七种药物的检测限在0.51 - 1.1 ng/mL范围内，特定药物的灵敏度主要由受体 - 药物亲和力决定，与结合能无关。综合比较后，本方法总体表现优于先前报道的氨基糖苷类酶联免疫吸附测定法。这是首次报道核糖体蛋白S12对氨基糖苷类药物的识别机制，并建立用于检测猪肉样品中氨基糖苷类药物残留的生物发光法的研究。
