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基于特异性单克隆抗体和无定形碳纳米粒子的高灵敏侧向流免疫分析法用于牛奶中磺胺类药物的检测。

A highly sensitive lateral flow immunoassay based on a group-specific monoclonal antibody and amorphous carbon nanoparticles for detection of sulfonamides in milk.

机构信息

Henan Engineering Technology Research Center of Food Processing and Circulation Safety Control, College of Food Science and Technology, Henan Agricultural University, 63 Nongye Road, Zhengzhou, 450002, Henan, China.

College of Food and Drug, Luoyang Normal University, Luoyang, 471934, Henan, China.

出版信息

Mikrochim Acta. 2023 Apr 18;190(5):186. doi: 10.1007/s00604-023-05766-3.

Abstract

To meet high-throughput screening of the residues of sulfonamides (SAs) with high sensitivity toward sulfamethazine (SM2) in milk samples, a new highly sensitive lateral flow immunoassay (LFA) based on amorphous carbon nanoparticles (ACNs) was developed. First, a group-specific monoclonal antibody 10H7 (mAb 10H7) that could recognize 25 SAs with high sensitivity toward SM2 (IC value of 0.18 ng/mL) was prepared based on H1 as an immune hapten and H4 as a heterologous coating hapten. Then, mAb 10H7 was conjugated to ACNs as an immune probe for LFA development. Under the optimized conditions, the LFA could detect 25 SAs with the cut-off value toward SM2 of 2 ng/mL, which could meet the requirement for detection of SAs. In addition, the LFA developed was also used for screening SAs' residues in real milk samples, with results being consistent with HPLC-MS/MS. Thus, this LFA can be used as a high-throughput screening tool for detection of SAs.

摘要

为了满足对牛奶样品中磺胺类药物(SAs)残留进行高通量筛选的要求,并对磺胺甲噁唑(SM2)具有高灵敏度,开发了一种基于无定形碳纳米粒子(ACNs)的新型高灵敏度侧向流动免疫分析(LFA)。首先,基于 H1 作为免疫半抗原和 H4 作为异源包被半抗原,制备了一种能够识别 25 种 SAs 且对 SM2 具有高灵敏度的组特异性单克隆抗体 10H7(mAb 10H7)(IC 值为 0.18 ng/mL)。然后,将 mAb 10H7 偶联到 ACNs 上作为 LFA 开发的免疫探针。在优化条件下,LFA 可以检测到 25 种 SAs,SM2 的截断值为 2 ng/mL,这可以满足 SAs 检测的要求。此外,该 LFA 还用于筛选实际牛奶样品中的 SAs 残留,结果与 HPLC-MS/MS 一致。因此,该 LFA 可作为一种用于检测 SAs 的高通量筛选工具。

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