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抗独特型纳米抗体-碱性磷酸酶融合蛋白：用于谷物中伏马菌素B1检测的一步竞争性酶免疫测定法的开发。

Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: Development of a one-step competitive enzyme immunoassay for fumonisin B1 detection in cereal.

作者信息

Shu Mei, Xu Yang, Liu Xing, Li Yanping, He Qinghua, Tu Zhui, Fu Jinheng, Gee Shirley J, Hammock Bruce D

机构信息

State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China; Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China.

State Key Laboratory of Food Science and Technology, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China; Jiangxi-OAI Joint Research Institute, Nanchang University, No. 235 Nanjing East Road, Nanchang 330047, People's Republic of China.

出版信息

Anal Chim Acta. 2016 Jun 14;924:53-59. doi: 10.1016/j.aca.2016.03.053. Epub 2016 Apr 7.

DOI:10.1016/j.aca.2016.03.053
PMID:27181644
Abstract

A rapid and sensitive one-step competitive enzyme immunoassay for the detection of FB1 was developed. The anti-idiotypic nanobody-alkaline phosphatase (Ab2β-Nb-AP) was validated by the AP enzyme activity and the properties of bounding to anti-FB1-mAb (3F11) through colorimetric and chemiluminescence analyses. The 50% inhibitory concentration and the detection limit (LOD) of colorimetric enzyme-linked immunosorbent assay (ELISA) for FB1 were 2.69 and 0.35 ng mL(-1), respectively, with a linear range of 0.93-7.73 ng mL(-1). The LOD of the chemiluminescence ELISA (CLIA) was 0.12 ng mL(-1), and the IC50 was 0.89 ± 0.09 ng mL(-1) with a linear range of 0.29-2.68 ng mL(-1). Compared with LC-MS/MS, the results of this assay indicated the reliability of the Ab2β-Nb-AP fusion protein based one-step competitive immunoassay for monitoring FB1 contamination in cereals. The Ab2β-Nb-AP fusion proteins have the potential to replace chemically-coupled probes in competitive enzyme immunoassay systems.

摘要

开发了一种用于检测伏马毒素B1（FB1）的快速灵敏的一步竞争酶免疫测定法。通过比色法和化学发光分析，利用碱性磷酸酶（AP）的酶活性以及与抗FB1单克隆抗体（3F11）结合的特性，对抗独特型纳米抗体-碱性磷酸酶（Ab2β-Nb-AP）进行了验证。用于FB1的比色酶联免疫吸附测定（ELISA）的50%抑制浓度和检测限（LOD）分别为2.69和0.35 ng mL⁻¹，线性范围为0.93 - 7.73 ng mL⁻¹。化学发光ELISA（CLIA）的LOD为0.12 ng mL⁻¹，IC50为0.89 ± 0.09 ng mL⁻¹，线性范围为0.29 - 2.68 ng mL⁻¹。与液相色谱-串联质谱（LC-MS/MS）相比，该测定结果表明基于Ab2β-Nb-AP融合蛋白的一步竞争免疫测定法用于监测谷物中FB1污染的可靠性。Ab2β-Nb-AP融合蛋白有潜力在竞争酶免疫测定系统中取代化学偶联探针。

相似文献

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Anti-idiotypic nanobody-alkaline phosphatase fusion proteins: Development of a one-step competitive enzyme immunoassay for fumonisin B1 detection in cereal.

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[3]
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[4]
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[5]
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[6]
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[7]
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[8]
An Electrochemical Fiveplex Biochip Assay Based on Anti-Idiotypic Antibodies for Fast On-Site Detection of Bioterrorism Relevant Low Molecular Weight Toxins.

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[9]
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[10]
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