Development of a Phage-Displayed Nanobody-Based Competitive Immunoassay for the Sensitive Detection of Soybean Agglutinin

Soybean agglutinin (SBA) is a primary antinutritional factor in soybeans that can inhibit the growth of humans and mammals, disrupt the intestinal environment, and cause pathological changes. Therefore, detecting and monitoring SBA in foods is essential for safeguarding human health. In this paper,...

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Bibliographic Details
Published in:Foods 2024-06, Vol.13 (12), p.1893
Main Authors: Zhang, Menghan, Qiu, Yulou, You, Ajuan, Song, Siyi, Yang, Qin, Zhang, Biao, Fu, Xianshu, Ye, Zihong, Yu, Xiaoping
Format: Article
Language:English
Subjects:
Albumen
Antibodies
Antigens
Antinutrition factors
Biotin
Cloning
competitive immunoassay
Enzyme-linked immunosorbent assay
Enzymes
Immunoassay
Libraries
Nanobodies
phage-displayed nanobody
Phages
Proteins
soybean agglutinin
Soybeans
Streptavidin
streptavidin poly-HRP
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Summary:Soybean agglutinin (SBA) is a primary antinutritional factor in soybeans that can inhibit the growth of humans and mammals, disrupt the intestinal environment, and cause pathological changes. Therefore, detecting and monitoring SBA in foods is essential for safeguarding human health. In this paper, M13 phage-displayed nanobodies against SBA were isolated from a naive nanobody library. An M13 phage-displayed nanobody-based competitive enzyme-linked immunosorbent assay (P-cELISA) was then established for SBA analysis using biotinylated anti-M13 phage antibody (biotin-anti-M13) and streptavidin poly-HRP conjugate (SA-poly-HRP). The biotin-anti-M13@SA-poly-HRP probe can easily amplify the detection signal without the chemical modifications of phage-displayed nanobodies. The established P-cELISA presented a linear detection range of 0.56-250.23 ng/mL and a limit of detection (LOD) of 0.20 ng/mL, which was 12.6-fold more sensitive than the traditional phage-ELISA. Moreover, the developed method showed good specificity for SBA and acceptable recoveries (78.21-121.11%) in spiked wheat flour, albumen powder, and whole milk powder. This study proposes that P-cELISA based on biotin-anti-M13@SA-poly-HRP may provide a convenient and effective strategy for the sensitive detection of SBA.
ISSN:2304-8158
2304-8158
DOI:10.3390/foods13121893