Journal of Zhejiang University

Journal of Zhejiang University SCIENCE B 2014 Vol.15 No.2 P.165-172

Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp*

Author(s): Tossapon Wongtangprasert¹,Wirongrong Natakuathung¹,Umaporn Pimpitak²,Anumart Buakeaw²,Tanapat Palaga³,Kittinan Komolpis²,Nanthika Khongchareonporn²
Affiliation(s): 1. 1. Program in Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand more
Corresponding email(s): nanthika.k@chula.ac.th
Key Words: Oxytetracycline, Monoclonal antibody, Enzyme-linked immunosorbant assay (ELISA), Shrimp

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Tossapon Wongtangprasert, Wirongrong Natakuathung, Umaporn Pimpitak, Anumart Buakeaw, Tanapat Palaga, Kittinan Komolpis, Nanthika Khongchareonporn. Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp[J]. Journal of Zhejiang University Science B, 2014, 15(2): 165-172.

@article{title="Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp",
author="Tossapon Wongtangprasert, Wirongrong Natakuathung, Umaporn Pimpitak, Anumart Buakeaw, Tanapat Palaga, Kittinan Komolpis, Nanthika Khongchareonporn",
journal="Journal of Zhejiang University Science B",
volume="15",
number="2",
pages="165-172",
year="2014",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B1300181"
}

%0 Journal Article
%T Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp
%A Tossapon Wongtangprasert
%A Wirongrong Natakuathung
%A Umaporn Pimpitak
%A Anumart Buakeaw
%A Tanapat Palaga
%A Kittinan Komolpis
%A Nanthika Khongchareonporn
%J Journal of Zhejiang University SCIENCE B
%V 15
%N 2
%P 165-172
%@ 1673-1581
%D 2014
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B1300181

TY - JOUR
T1 - Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp
A1 - Tossapon Wongtangprasert
A1 - Wirongrong Natakuathung
A1 - Umaporn Pimpitak
A1 - Anumart Buakeaw
A1 - Tanapat Palaga
A1 - Kittinan Komolpis
A1 - Nanthika Khongchareonporn
J0 - Journal of Zhejiang University Science B
VL - 15
IS - 2
SP - 165
EP - 172
%@ 1673-1581
Y1 - 2014
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B1300181

Abstract
Chinese Summary
Academic Network
Reviewer Comment

Abstract: A novel monoclonal antibody (MAb) against oxytetracycline (OTC) was generated and characterized. The MAb was used in the development of an enzyme-linked immunosorbant assay (ELISA)-based detection system. An OTC-bovine serum albumin (BSA) conjugate was prepared and used in the immunization of mice. A conventional somatic cell fusion technique was used to generate MAb-secreting hybridomas denoted 2-4F, 7-3G, and 11-11A. An indirect competitive ELISA (icELISA) was applied to measure the sensitivity and specificity of each MAb in terms of its 50% inhibitory concentration (IC₅₀) and percentage of cross-reactivity, respectively. MAb 2-4F exhibited the highest sensitivity, with an IC₅₀ of 7.01 ng/ml. This MAb showed strong cross-reactivity to rolitetracycline, but no cross-reactivity to other unrelated antibiotics. When MAb 2-4F was used to detect OTC from shrimp samples, the recoveries were in the range of 82%–118% for an intra-assay and 96%–113% for an inter-assay. The coefficients of variation of the assays were 3.9%–13.9% and 5.5%–14.9%, respectively.

抗土霉素单克隆抗体的制备及其在虾中土霉素残留的检测应用

研究目的：抗土霉素单克隆抗体的制备和特性分析，并用于间接竞争酶联免疫吸附测定法（icELISA）分析虾样品中土霉素的残留。
创新要点：本研究建立分泌抗土霉素单克隆抗体的杂交瘤细胞株，筛选出对虾中土霉素残留检测具有较高灵敏度的单克隆抗体2-4F。
研究方法：用细胞杂交技术使土霉素-牛血清白蛋白偶联物免疫的BALC/c雌性小鼠的脾细胞与骨髓瘤细胞融合，建立三株分泌抗土霉素单克隆抗体的杂交瘤细胞株（2-4F、7-3G和11-11A），并制备它们的单克隆抗体。通过icELISA法分析单克隆抗体对土霉素的半抑制质量浓度（IC₅₀）和交叉反应率，明确其灵敏度和特异性。
重要结论：实验结果发现单克隆抗体2-4F具有最高的灵敏度，对土霉素的IC₅₀为7.01 ng/ml，且该单抗特异性强，仅与罗利环素之间有强烈的交叉反应，而与非相关抗生素不发生交叉反应。当利用单克隆抗体2-4F检测虾样品中的土霉素含量时，其批内和批间回收率分别为82%~118%和96%~113%，变异系数分别为3.9%~13.9%和5.5%~14.9%。因此，单克隆抗体2-4F可用于虾产品中土霉素残留分析试剂盒的开发。

关键词：土霉素；单克隆抗体；酶联免疫吸附测定法（ELISA）；虾