数字农科院2.0

Genetically engineered integrated aflatoxin B1 and deoxynivalenol bispecific nanobody as surrogate antigens for constructed time-resolved immunoassay dual detection methods

文献类型： 外文期刊

作者： 袁贝;;李志强;;李培武;;张奇;;杨青青;;唐晓倩

关键词： Bispecific nanobody Immunochromatographic assay Mycotoxin detection One-step coupling Surrogate antigen

期刊名称： BIOSENSORS & BIOELECTRONICS

ISSN： 0956-5663

年卷期： 2025 年

页码：

收录情况： SCIE（2025版） ; ; EI（2025版）

摘要： There is a phenomenon of combined contamination of fungal toxins, of which aflatoxin B1 (AFB1) is the most toxic, and deoxynivalenol (DON) contamination is common. The use of antigens for double or multiple testing of mycotoxins is easy to cause environmental pollution, and surrogate antigens have become necessary. The small molecule and susceptibility to genetic modification of nanobodies can be used to develop alternative antigens for mycotoxins. In this study, using the nanobody gene sequences of the heavy chain recognition regions of anti-aflatoxin and deoxynivalenol monoclonal antibodies, recombinant plasmids were successfully constructed by one-step cloning, and low-temperature-induced bispecific nanobodies against AFB1-DON were obtained, which can be used as alternative antigens to reduce the pollution of the environment from mycotoxin detection. Enzyme-linked immunosorbent assay validated the bispecific nanobody, and the semi-inhibitory concentration (IC50) of the bispecific nanobody were 0.47 μg/L and 149 μg/L for AFB1 and DON, respectively. Finally, a time-resolved fluorescent dual-detection test strip was constructed by this bispecific nanobody as a surrogate antigen for AFB1 and DON, which was capable of detecting AFB1 and DON at the same time, and the limits of detection (LOD) for the two toxins were 0.0254 μg/L and 21.4 μg/L, respectively. This method has satisfactory sensitivity and does not require antigen, which reduces the toxicity of using antigen.

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