Preparation of Universal Hapten Immunochromatographic Strip and Its Application in Imidocarb

doi:10.15933/j.cnki.1004-3268.2024.09.012

Journal of Henan Agricultural Sciences ›› 2024, Vol. 53 ›› Issue (9): 117-125.DOI: 10.15933/j.cnki.1004-3268.2024.09.012

• Animal Husbandry and Veterinary Medicine • Previous Articles Next Articles

Preparation of Universal Hapten Immunochromatographic Strip and Its Application in Imidocarb

XING Yunrui¹，SUN Yaning¹，HU Xiaofei¹，FAN Lu¹，CHEN Linlin²，ZHANG Gaiping¹

（1.Institute for Animal Health(Key Laboratory of Animal Immunology),Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;2.College of Food Science and Technology，Henan Agricultural University,Zhengzhou 450002,China）

Received:2023-10-16 Published:2024-09-15 Online:2024-11-04

一种通用型半抗原免疫层析试纸的制备及在双咪苯脲上的应用

邢云瑞¹，孙亚宁¹，胡骁飞¹，范璐¹，陈琳琳²，张改平¹

（1.河南省农业科学院动物疫病防控研究所（河南省农业科学院动物免疫学重点实验室），河南郑州 450002；2.河南农业大学食品科学技术学院，河南郑州 450002）

通讯作者: 张改平（1960-），男，河南内黄人，研究员，博士，主要从事动物免疫学及动物病毒分子致病机制研究。E-mail：zhanggaip@126.com
作者简介:邢云瑞（1990-），女，河南舞阳人，助理研究员，硕士，主要从事免疫学检测技术研究。E-mail：xyrui1201@163.com
基金资助:
河南省农业科学院自主创新项目（2023ZC086）；河南省重点研发与推广专项（科技攻关）（222102110413）；河南省农业科学院优秀青年科技基金项目（2022YQ25）；国家重点研发计划项目（2018YFC1602903）

Abstract

Abstract: In order to improve the selection efficiency of monoclonal antibodies and lower the preparation threshold of immunochromatographic strip,a universal hapten immunochromatographic strip was established by using biotin‑labeled artificial antigen as a probe and avidin in a test line,and applied for selecting monoclonal antibodies against imidocarb(IM)and detecting IM residues.The results showed that IM artificial antigen and biotin probe were successfully prepared by 1‑(3‑dimethylaminopropyl)‑3‑ethylcarbodiimide(EDC)method.Subsequently,monoclonal antibodies were prepared by immunizing mice and hybridoma cells technology,and the universal immunochromatographic strip was established.Using the strip,a monoclonal antibody cell line against IM with a titer of 1:32 000 was successfully selected,and its sensitivity was up to 6.25 ng/mL.Based on the excellent monoclonal antibody and the universal strip,an immunochromatographic test strip for the detection of IM was established,and its sensitivity was 7 ng/mL in the milk.The immunochromatographic strip and its probe preparation method can be used in general hapten antibody screening,and a hapten immunochromatographic strip test can be established without professional research and development platform,which provides a more efficient method for antibody screening and a new technology for the development of immunochromatographic strip.

Key words: Hapten, Universal immunochromatographic strip, Imidocarb, Antibody selection, Residue detection

摘要： 为了提高单克隆抗体筛选效率，降低免疫层析检测方法建立门槛，利用生物素标记人工抗原为探针，以亲和素固定于硝酸纤维素膜上为检测线，建立了通用型半抗原免疫层析试纸，并将其应用于双咪苯脲（IM）单克隆抗体的筛选及残留检测。结果表明，利用碳二亚胺法成功制备了IM人工抗原及生物素探针；通过免疫小鼠及杂交瘤细胞技术制备单克隆抗体；应用试纸筛选，成功获得IM单克隆抗体细胞株1株，其效价为1∶32 000，灵敏度为6.25 ng/mL。基于该单抗及通用型半抗原免疫层析试纸建立了IM免疫层析检测试纸，检测牛奶中IM的灵敏度为7 ng/mL。该试纸及其探针制备方法可以通用于半抗原抗体筛选，为抗体筛选提供了一种更高效的方法；无需免疫层析试纸研发平台即可建立半抗原免疫层析检测方法，为免疫层析试纸的研发提供了新的技术手段。

关键词: 半抗原, 通用型免疫层析试纸, 双咪苯脲, 抗体筛选, 残留检测

CLC Number:

R446.6

XING Yunrui, SUN Yaning, HU Xiaofei, FAN Lu, CHEN Linlin, ZHANG Gaiping. Preparation of Universal Hapten Immunochromatographic Strip and Its Application in Imidocarb[J]. Journal of Henan Agricultural Sciences, 2024, 53(9): 117-125.

邢云瑞, 孙亚宁, 胡骁飞, 范璐, 陈琳琳, 张改平. 一种通用型半抗原免疫层析试纸的制备及在双咪苯脲上的应用[J]. 河南农业科学, 2024, 53(9): 117-125.

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Preparation of Universal Hapten Immunochromatographic Strip and Its Application in Imidocarb

一种通用型半抗原免疫层析试纸的制备及在双咪苯脲上的应用

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