食品与发酵工业 >

2025 , Vol. 51 >Issue 17: 335 - 344

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.042027

分析与检测

五氯酚酸钠单克隆抗体的制备及胶体金免疫层析法的建立

  • 刘波 ,
  • 华彦涛 ,
  • 黄皓璋 ,
  • 区惠珍 ,
  • 李斌 ,
  • 袁利鹏 ,
  • 刘诗文
展开
  • 1(广东农工商职业技术学院 热带农林学院,广东 广州,511365)
    2(广州万联生物科技有限公司,广东 广州,510663)
    3(吕梁学院 生物与食品工程系,山西 吕梁,033001)
第一作者:硕士,副教授 (袁利鹏教授为通信作者, E-mail:543753927@qq.com)

收稿日期: 2025-01-02

  修回日期: 2025-04-15

  网络出版日期: 2025-09-29

基金资助

广东省教育厅普通高校重点科研平台项目(2024KCXTD082,2019GGCZX010);2022年广东省攀登计划项目(pdjh2022b0914);广州市增城高层次人才项目;广东省普通高校青年创新人才类项目(2022KQNCX211)

收起

Preparation of monoclonal antibody and establishment of colloidal gold immunochromatography assay for sodium pentachlorophenolate

  • LIU Bo ,
  • HUA Yantao ,
  • HUANG Haozhang ,
  • OU Huizhen ,
  • LI Bin ,
  • YUAN Lipeng ,
  • LIU Shiwen
Expand
  • 1(Tropical agriculture and Forestry College, Guangdong AIB Polytechnic College, Guangzhou 511365, China)
    2(Guangzhou Wanlian Biotechnology Co.Ltd., Guangzhou 510663, China)
    3(Department of Biological and Food Engineering, Lyuliang University, Lyuliang 033001, China)

Received date: 2025-01-02

  Revised date: 2025-04-15

  Online published: 2025-09-29

Fold

摘要

为了实现更快速、灵敏、准确检测食品中五氯酚酸钠(sodium pentachlorophenoxide,PCP-Na)的残留,以PCP-Na的结构为基础,通过保留其苯环上的酚羟基设计合成了半抗原,制备了特异性识别PCP-Na的单克隆抗体,并建立一种快速检测动物性食品中PCP-Na的胶体金免疫层析法。通过优化标记体系、抗原包被质量浓度和样品前处理等关键参数,在最优条件下,所制备的抗PCP-Na单克隆抗体的半数抑制浓度为0.87 μg/L,所建立的胶体金免疫层析法对PCP-Na的标准品检出限、样品检出限分别为0.8 μg/kg、1 μg/kg,具有灵敏、准确、快速等特点。该法可应用于鸡肉、猪肉、罗非鱼、生蚝等动物性食品中PCP-Na的大批量快速筛选。

关键词： PCP-Na; 单克隆抗体; 胶体金免疫层析法; 动物性食品; 快速检测

本文引用格式

刘波 , 华彦涛 , 黄皓璋 , 区惠珍 , 李斌 , 袁利鹏 , 刘诗文 . 五氯酚酸钠单克隆抗体的制备及胶体金免疫层析法的建立[J]. 食品与发酵工业, 2025 , 51(17) : 335 -344 . DOI: 10.13995/j.cnki.11-1802/ts.042027

Abstract

To achieve a faster, sensitive, and accurate detection of sodium pentachlorophenolate (PCP-Na) residues in food, based on the structure of PCP-Na, a hapten was designed and synthesised by retaining the phenolic hydroxyl group on the benzene ring, a monoclonal antibody specifically recognizing PCP-Na was prepared, and a colloidal gold immunochromatographic method for the rapid detection of PCP-Na in animal food was established.By the key parameters such as labelling system, antigen coating concentration and sample preparation, the half-inhibition concentration (IC50) of the prepared anti-PCP-Na monoclonal antibody was 0.87 μg/L under optimal conditions.The established colloidal gold immunochromatography method had the characteristics of sensitive, accurate, and rapid.The detection limits of PCP-Na and samples were 0.8 and 1 μg/kg, respectively.This method can be applied to the large-scale rapid screening of PCP-Na in animal foods such as chicken, pork, tiplapia fish, and oyster.

Key words： sodium pentachlorophenolate; monoclonal antibody; colloidal gold immunochromatography; animal food; rapid detection

参考文献

[1] 何正和, 陆井莲, 姜颖慧, 等.水产品中五氯苯酚及其钠盐检测技术研究进展[J].食品安全导刊, 2023(9):13-15.
HE Z H, LU J L, JIANG Y H, et al.Research progress in detection technology of pentachlorophenol and its sodium salt in aquatic products[J].China Food Safety Magazine, 2023(9):13-15.
[2] ZHANG W X, TANG Y, HAN Y, et al.Immunotoxicity of pentachlorophenol to a marine bivalve species and potential toxification mechanisms underpinning[J].Journal of Hazardous Materials, 2022, 439:129681.
[3] YANG L H, ZHA J M, WANG Z J.Pentachlorophenol affected both reproductive and interrenal systems:In silico and in vivo evidence[J].Chemosphere, 2017, 166:174-183.
[4] 陈彦宏, 黄松, 陈穗, 等.五氯酚及其钠盐的样品前处理和分析检测技术研究进展[J].食品安全质量检测学报, 2019, 10(14):4465-4473.
CHEN Y H, HUANG S, CHEN S, et al.Research progress of sample pretreatment and analytical techniques for determination of pentachlorophenol and sodium pentachlorophenate[J].Journal of Food Safety & Quality, 2019, 10(14):4465-4473.
[5] ZHANG X N, KANG H, PENG L Z, et al.Pentachlorophenol inhibits CatSper function to compromise progesterone's action on human sperm[J].Chemosphere, 2020, 259:127493.
[6] 阮鑫, 陆春波, 屈健.畜禽产品中五氯酚酸钠残留危害与分析[J].中国畜禽种业, 2022, 18(1):54-56.
RUAN X, LU C B, QU J.Harm and analysis of sodium pentachlorophenol residue in livestock and poultry products[J].The Chinese Livestock and Poultry Breeding, 2022, 18(1):54-56.
[7] 张秀芹, 王玉晶, 李胤楠, 等.动物源产品中五氯酚来源探讨及危害分析[J].食品安全质量检测学报, 2021, 12(19):7840-7845.
ZHANG X Q, WANG Y J, LI Y N, et al.Discussion on source and hazard analysis of pentachlorophenol in animal-derived products[J].Journal of Food Safety & Quality, 2021, 12(19):7840-7845.
[8] MAHESHWARI N, KHAN A A, MAHMOOD R, et al.Pentachlorophenol-induced hemotoxicity diminishes antioxidant potential and oxidizes proteins, thiols, and lipids in rat blood:An in vivo study[J].Heliyon, 2023, 9(5):e16240.
[9] HUANG S R, CHANG J L, XU Z L, et al.Robust and bioaffinity-enhanced nanocarrier based immunochromatographic assay with simplified sample preparation for pentachlorophenol sodium in animal tissues[J].Talanta, 2024, 267:125190.
[10] 李容. 固相萃取-气相色谱质谱法测定鸡肉中五氯酚钠的研究[J].食品安全导刊, 2019(30):105.
LI R.Determination of sodium pentachlorophenol in chicken by solid phase extraction-gas chromatography-mass spectrometry[J].China Food Safety Magazine, 2019(30):105.
[11] 潘明迪. 液相色谱-三重四极杆串联质谱结合QuEChERS测定牛百叶中五氯酚酸钠的含量[J].食品安全导刊, 2023(12):45-47;53.
PAN M D.Determination of sodium pentachlorophenate in bovine louver by liquid chromatography-triple quadrupole tandem mass spectrometry combined with QuEChERS[J].China Food Safety Magazine, 2023(12):45-47;53.
[12] 张美, 朱莉莎, 徐飘飘, 等.高效液相色谱—串联质谱法检测鸡蛋中五氯酚钠药物残留研究[J].现代畜牧兽医, 2023(3):19-22.
ZHANG M, ZHU L S, XU P P, et al.Determination of pentachlorophenol sodium residues in eggs by high performance liquid chromatography tandem mass spectrometry[J].Modern Journal of Animal Husbandry and Veterinary Medicine, 2023(3):19-22.
[13] DONG S, GUAN L J, ZHAO Y, et al.A dual-signal immunochromatographic assay using quantum dots and polydopamine coated gold nanoparticles for detection of sodium pentachlorophenate in animal-derived food[J].Microchemical Journal, 2022, 183:108070.
[14] DONG S, SHI Q Y, GUAN L J, et al.Preparation of monoclonal antibody and establishment of indirect competitive chemiluminescence enzyme immunoassay for sodium pentachlorophenolate[J].Microchemical Journal, 2024, 196:109674.
[15] HUANG S R, CHEN Z Y, WANG Y C, et al.Post-synthetic modification fluorescence UiO-66-Eu immunochromatography for high-performance detection of sodium pentachlorophenoate[J].Journal of Hazardous Materials, 2024, 480:135824.
[16] 龚蕾, 周陶鸿, 彭青枝, 等.免疫层析法快速检测动物源性食品中五氯酚酸钠含量[J].食品安全质量检测学报, 2022, 13(3):888-893.
GONG L, ZHOU T H, PENG Q Z, et al.Rapid determination of sodium pentachlorophenolate in animal-derived foods by immunochromatography[J].Journal of Food Safety & Quality, 2022, 13(3):888-893.
[17] 罗晓琴, 韩深, 吕艳秋, 等.五氯酚钠胶体金快速检测试纸条的研制[J].安徽农业科学, 2016, 44(25):55-57.
LUO X Q, HAN S, LU Y Q, et al.Study on gold immunochromatography assay for rapid detection of pentachlorophenol-Na[J].Journal of Anhui Agricultural Sciences, 2016, 44(25):55-57.
[18] 余宇燕, 张淑玲, 邹艳辉, 等.五氯酚免疫层析检测试纸条的研究[J].分析测试学报, 2014, 33(7):830-834.
YU Y Y, ZHANG S L, ZOU Y H, et al.Preparation of colloidal gold immunochromatography strip for detection of pentachlorophenol[J].Journal of Instrumental Analysis, 2014, 33(7):830-834.
[19] ZHANG X Y, BAI Y C, TANG Q Q, et al.Development of epitopephore-based rational hapten design strategy:A combination of theoretical evidence and experimental validation[J].Journal of Hazardous Materials, 2023, 445:130615.
[20] ZHANG L W, WANG J C, WANG Y T, et al.Preparation of monoclonal antibodies recognizing pharmacologically active metabolites of metamizole based on rational hapten design and their application in the detection of animal-derived food[J].Talanta, 2024, 280:126753.
[21] LIANG J X, LIU Z W, FANG Y L, et al.Two kinds of lateral flow immunoassays based on multifunctional magnetic Prussian blue nanoenzyme and colloidal gold for the detection of 38 β-agonists in swine urine and pork[J].Food Chemistry, 2023, 417:135897.
[22] KAKKAR S, GUPTA P, SINGH YADAV S P, et al.Lateral flow assays:Progress and evolution of recent trends in point-of-care applications[J].Materials Today Bio, 2024, 28:101188.
Options
文章导航

/

〈 〉

技术支持：北京玛格泰克科技发展有限公司