A novel yeast-based biosensor for the quick determination of Deoxynivalenol.
Anal Chim Acta
; 1315: 342760, 2024 Aug 01.
Article
em En
| MEDLINE
| ID: mdl-38879206
ABSTRACT
Mycotoxins are commonly found in food materials and severely threaten human health. Antibodies play a key role as a part of immunological techniques in detecting mycotoxins. Therefore, highly specific antibodies and detection techniques against mycotoxins need to be developed for advancements in medical research. In this study, we presented a novel strategy for quickly screening highly specific antigen-binding fragment (Fab) antibodies based on yeast surface display (YSD) and detecting small-molecule compounds based on a YSD biosensor. We constructed a yeast surface display Deoxynivalenol (DON)-Fab library with 105 cfu/mL with a galactose-inducible bidirectional promoter. By conducting efficient magnetic-activated cell sorting and fluorescence-activated cell sorting (MACS/FACS), four kinds of DON-selective yeasts were screened. As Fab@YSD C4# showed high sensitivity, we used it to build a one-pot Fab@YSD chemiluminescence biosensor with DON-BSA@Biotin and Streptavidin-alkaline phosphatase (SA-ALP). This method showed a low operational threshold (LOD = 0.166 pg/mL) and a high population range (linear range = 0.001-132.111 ng/mL) within 40 min, which facilitated the detection of DON with high specificity and better recovery in real samples (wheat, corn, flour, and cornmeal). Our results suggested that the Fab@YSD chemiluminescence biosensor is an inexpensive, reproducible, user-friendly, and sensitive method for detecting DON and may be used to quickly detect other small-molecule contaminants in food items.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Tricotecenos
/
Técnicas Biossensoriais
Idioma:
En
Revista:
Anal Chim Acta
Ano de publicação:
2024
Tipo de documento:
Article