微囊藻毒素纳米抗体的制备及其间接竞争酶联免疫分析方法的建立
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(华南农业大学食品学院 广东省食品质量与安全重点实验室畜产品精准加工与安全控制技术国家地方联合工程研究中心(广东) 广州 510642)

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司睿(1993—),男,硕士生

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“十三五”国家重点研发计划重点专项(2019 YFE0116600);国家自然科学基金项目(319721 57);广东省重点领域研发计划项目(2019B 020211002);广州市科技计划项目(202102080 615);广东省高等学校珠江学者岗位计划项目(2017)


Preparation of Microcystin Nanobody and Establishment of Indirect Competitive Enzyme-linked Immunosorbent Assay for Microcystin
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(Guangdong Provincial Key Laboratory of Food Quality and Safety, National-Local Joint Engineering Research Center for Processing and Safety Control of Livestock and Poultry Products (Guangdong), College of Food Science, South China Agricultural University, Guangzhou 510642)

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    摘要:

    微囊藻毒素(MC)作为肝毒性最强的藻毒素之一,在受污染的水体中时常检出,严重威胁着人畜饮水安全,加强其检测十分重要。在前期研究中,已成功构建抗MC-LR噬菌体展示纳米抗体文库。本研究以MC-LR-OVA作为包被原,经4轮固相淘筛,获得纳米抗体M110。该抗体可识别MC-LR,且具有良好的稳定性。基于纳米抗体M110建立了检测MC-LR的间接竞争酶联免疫分析法(ic-ELISA),检测的半抑制浓度IC50值为3.55 ng/mL,检测限0.65 ng/mL,线性范围1.28~9.88 ng/mL。与UPLC-MS/MS方法检测结果相关系数达0.998,提示本方法可用于水体中微囊藻毒素残留的检测。

    Abstract:

    Microcysin (MC), as one of the most toxic hepatoxins, is often detected in polluted water bodies, which seriously threatens the safety of drinking water for humans and animals. It is very important to develop its rapid detection methods. In the previous study, an anti-MC-LR phage display nanobody library was successfully constructed. In this study, using MC-LR-OVA as the coating antigen, the anti-MC-LR nanobody M110 was selected by four rounds solid phase biopanning. M110 could recognize MC-LR with good stability. Further, an indirect competiitive enzyme-linked immunosorbent assay (ic-ELISA) for the detection of MC-LR was established, with the half-maximal inhibitory concentration (IC50) of 3.55 ng/mL. The detection limit was 0.65 ng/mL and the linear range was 1.28-9.88 ng/mL. The correlation coefficient of ic-ELISA and UPLC-MS/MS was 0.998, indicating the M110-based ic-ELISA method has high accuracy and can be used for the actual detection of MC-LR residues in water sample.

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司睿,吴广培,王锋,雷红涛,王弘.微囊藻毒素纳米抗体的制备及其间接竞争酶联免疫分析方法的建立[J].中国食品学报,2022,22(5):332-337

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  • 收稿日期:2021-05-08
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  • 在线发布日期: 2022-06-15
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