生产高亲和力单克隆抗体并建立用于动物组织中庆大霉素残留的间接竞争化学发光酶免疫分析方法。

Production of high affinity monoclonal antibody and development of indirect competitive chemiluminescence enzyme immunoassay for gentamicin residue in animal tissues.

机构信息

Key Laboratory of Agricultural Products Processing and Quality Control of Nanchang City/College of Food Science and Engineering, Jiangxi Agricultural University, Nanchang 330045, China.

Guangdong Testing Institute of Product Quality Supervision, Shunde 528300, China.

出版信息

Food Chem. 2023 Jan 30;400:134067. doi: 10.1016/j.foodchem.2022.134067. Epub 2022 Sep 5.

DOI:10.1016/j.foodchem.2022.134067

PMID:36084594

Abstract

To determine gentamicin residues in animal tissues, a monoclonal antibody (Mab) was produced and a sensitive indirect competitive chemiluminescent enzyme immunoassay (icCLEIA) was developed. At first, gentamicin was conjugated with bovine serum albumin as immunogens which were used to immunize BALB/c mice. Then, an anti-gentamicin Mab was prepared by hybridoma technology. Finally, a sensitive icCLEIA was developed with an 50% inhibition concentration (IC) of 0.067 ng/mL for gentamicin. The limit of detection of the icCLEIA was 0.002 ng/mL. The cross reactivity of the Mab with structural analogues were<0.01%. The recoveries of gentamicin ranged from 80 to 101% and coefficient of variation was <6.4% in pork and fish samples. Samples were detected by UPLC-MS/MS for evaluating reliability of the icCLEIA. The results suggested that the prepared anti-gentamicin Mab can be used for rapid and convenient immunoassays to detect gentamicin residues in animal tissues.

摘要

为了检测动物组织中的庆大霉素残留，制备了一种单克隆抗体（Mab），并开发了一种灵敏的间接竞争化学发光酶免疫分析（icCLEIA）方法。首先，庆大霉素与牛血清白蛋白偶联作为免疫原，用于免疫 BALB/c 小鼠。然后，通过杂交瘤技术制备抗庆大霉素 Mab。最后，建立了一种灵敏的 icCLEIA，庆大霉素的半数抑制浓度（IC）为 0.067ng/mL。icCLEIA 的检测限为 0.002ng/mL。该 Mab 与结构类似物的交叉反应性<0.01%。在猪肉和鱼肉样品中，庆大霉素的回收率为 80%至 101%，变异系数<6.4%。采用 UPLC-MS/MS 检测样品，以评估 icCLEIA 的可靠性。结果表明，所制备的抗庆大霉素 Mab 可用于快速、方便地免疫分析检测动物组织中的庆大霉素残留。