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• W4387345479 endingPage "132701" @default.
• W4387345479 startingPage "132701" @default.
• W4387345479 abstract "Mycotoxins produced by fungi can contaminate various foods and pose significant health risks. Ensuring food safety demands rapid, highly sensitive analytical techniques. One-step Bioluminescent Enzyme Immunoassays (BLEIAs) employing nanobody-nanoluciferase fusion proteins have recently garnered attention for operational simplicity and heightened sensitivity. Nevertheless, fixed nanobody:nanoluciferase ratios in fusion proteins restrict the customization and sensitivity of traditional BLEIAs. In this study, we present a Scaffold Assembly-based BLEIA (SA-BLEIA) that overcomes these limitations through the programmable conjugation of nanobodies and luciferases onto 60-meric protein nanoscaffolds using SpyTag/SpyCatcher linkages. These nanoscaffolds facilitate the adjustable coupling of anti-aflatoxin B1 and anti-ochratoxin A nanobodies with luciferases, optimizing nanobody/luciferase ratios and diversifying specificities. Compared to conventional methods, SA-BLEIA demonstrates considerably elevated sensitivity for detecting both toxins. The elevated local concentration of luciferase significantly amplifies bioluminescence intensity, permitting reduced substrate consumption and cost-effective detection. The usage of dual-nanobody conjugates facilitates the quantification or simultaneous detection of both mycotoxins in a single test with shared reagents. The assay exhibits exceptional recovery rates in spiked cereal samples, strongly correlating with outcomes from commercial ELISA kits. Overall, this adaptable, highly sensitive, cost-effective, and multiplexed immunoassay underscores the potential of tunable scaffold assembly as a promising avenue for advancing bioanalytical diagnostic tools." @default.
• W4387345479 created "2023-10-05" @default.
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• W4387345479 date "2024-01-01" @default.
• W4387345479 modified "2023-10-17" @default.
• W4387345479 title "Protein Nanoscaffold Enables Programmable Nanobody-Luciferase Immunoassembly for Sensitive and Simultaneous Detection of Aflatoxin B1 and Ochratoxin A" @default.
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• W4387345479 doi "https://doi.org/10.1016/j.jhazmat.2023.132701" @default.
• W4387345479 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/37839380" @default.
• W4387345479 hasPublicationYear "2024" @default.
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