RT Journal Article
T1 Biotechnology-enhanced immunoassay for accurate determination of HT-2 toxin in edible insect samples
A1 Sancho-García, Raúl
A1 Navarro Villoslada, Fernando
A1 Prádanas González, Fernando
A1 Arola, Henri O.
A1 Glahn Martínez, Ana Bettina
A1 Nevanen, Tarja K.
A1 Benito Peña, María Elena
AB Consumption of edible insects is common in non-Western countries of Africa, Asia, Oceania, and Latin America. However, their consumption has significantly increased in Europe in recent years thanks to their remarkable nutritional properties. Edible insects provide a valuable source of high-quality proteins, fats, minerals, and vitamins. Nevertheless, the absence of global regulatory guidelines poses a risk associated with their consumption due to the potential presence of pathogens and contaminants, such as mycotoxins, which are toxic compounds produced by fungi and represent a major threat to food and feed safety. Our approach integrates advanced nanobiotechnology to develop fluorescent antibodies by conjugating a recombinant superfolder green fluorescent protein (sfGFP) with a single-chain antibody (scFv). This fusion allows for precise detection of the immune complex formed between the HT-2 toxin and a biotinylated anti-HT-2 antibody. Additionally, we employed advanced computational tools, including AlphaFold and MOE, to deepen our understanding of the binding interactions present in the immune complex, confirming the strong interaction between the Fab/HT-2 toxin immunocomplex and the scFv antibody fragment, in contrast to the weaker binding observed with the Fab/T-2 toxin and the scFv. The method demonstrates high sensitivity, with an EC50 of 10.3 ± 0.6 ng mL−1, a dynamic range of 3.4 ± 0.1 to 31 ± 3 ng mL−1, a limit of detection of 0.43 ng mL−1, and a limit of quantification of 1.2 ng mL−1 in buffer solution. The assay exhibited excellent precision, with a reproducibility of 4% and no cross-reactivity with other mycotoxins. Application to contaminated cricket flour yielded recoveries between 91 and 133%, with coefficients of variation from 6 to 13%. These results indicate that the developed immunoassay is highly sensitive, selective, and reliable for detecting HT-2 toxin in food matrices, providing a promising tool for mycotoxin screening in food safety.
PB Springer Nature
SN 0026-3672
SN 1436-5073
YR 2025
FD 2025-04-16
LK https://hdl.handle.net/20.500.14352/124187
UL https://hdl.handle.net/20.500.14352/124187
LA eng
NO Sancho-García, R., Navarro-Villoslada, F., Pradanas-González, F. et al. Biotechnology-enhanced immunoassay for accurate determination of HT-2 toxin in edible insect samples. Microchim Acta 192, 301 (2025)
DS Docta Complutense
RD 17 dic 2025