当前位置: X-MOL 学术Food Agric. Immunol. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Development of a single-chain variable fragment-alkaline phosphatase fusion protein-based direct competitive enzyme-linked immunosorbent assay for furaltadone metabolite in ctenopharyngodon idellus
Food and Agricultural Immunology ( IF 3 ) Pub Date : 2021-08-01 , DOI: 10.1080/09540105.2021.1957781
Xinzhu Chen 1 , Huaying Du 1 , Yanping Hong 1 , Yubo Wang 1 , Mei Shu 1 , Dan Wang 1 , Peng Dai 1 , Weijie Deng 1 , Jianhua Xiong 1 , Wuying Yang 1
Affiliation  

ABSTRACT

To determine furaltadone metabolites 5-methylmorpholino-3-amino-2-oxazolidinone (AMOZ) residual levels in ctenopharyngodon idellus, direct competitive enzyme-linked immunosorbent assay (dcELISA) was developed based on a single-chain variable fragment (scFv)-alkaline phosphatase (AP) fusion protein. Firstly, VH and VL gene sequences were cloned from hybridoma cell lines secreting monoclonal antibody against AMOZ derivative, and then the productive VH and VL genes were assembled to a scFv gene and engineered to produce scFv-AP fusion protein. The fusion protein was further identified as a bifunctional reagent for immunoassay, then a sensitive one-step dcELISA against AMOZ derivative was developed with a half-maximal inhibitory concentration (IC50) and limit of detection (LOD) of 10.62 and 4.83 ng/mL, respectively. The recovery values of AMOZ from spiked fish samples determined by dcELISA were in 71.31%∼82.88%. The results suggested that the prepared scFv-AP fusion protein in this work can be used for rapid and convenient immunoassays to detect AMOZ residues in fish samples.



中文翻译:

基于单链可变片段-碱性磷酸酶融合蛋白的直接竞争性酶联免疫吸附测定法检测草鱼中呋喃他酮代谢物的研究

摘要

以确定代谢物呋喃他酮5-甲基吗啉代-3-氨基甲基-2-恶唑烷酮(AMOZ)中残留水平草鱼,直接竞争酶联免疫吸附测定(dcELISA)的基础上的单链可变片段(scFv) -碱性磷酸酶被开发(AP) 融合蛋白。首先,从分泌抗AMOZ衍生物单克隆抗体的杂交瘤细胞系中克隆V HVL基因序列,然后生产V HVL基因被组装成一个 scFv 基因并被改造以产生 scFv-AP 融合蛋白。融合蛋白被进一步鉴定为免疫测定的双功能试剂,然后开发了针对 AMOZ 衍生物的灵敏一步 dcELISA,半数抑制浓度 (IC 50 ) 和检测限 (LOD) 分别为 10.62 和 4.83 ng/mL , 分别。dcELISA测定的加标鱼样品中AMOZ的回收率为71.31%~82.88%。结果表明,本工作制备的scFv-AP融合蛋白可用于快速方便的免疫分析,检测鱼类样品中的AMOZ残留。

更新日期:2021-08-02
down
wechat
bug