Production and Characterization of Biotinylated Anti-fenitrothion Nanobodies and Development of Sensitive Fluoroimmunoassay.
biotinylation
carbon dots
fenitrothion
fluoroimmunoassay
nanobody
Journal
Journal of agricultural and food chemistry
ISSN: 1520-5118
Titre abrégé: J Agric Food Chem
Pays: United States
ID NLM: 0374755
Informations de publication
Date de publication:
06 Apr 2022
06 Apr 2022
Historique:
pubmed:
26
3
2022
medline:
8
4
2022
entrez:
25
3
2022
Statut:
ppublish
Résumé
A simple and sensitive fluoroimmunoassay (FIA) based on a heavy-chain antibody (VHH) for rapid detection of fenitrothion was developed. A VHH library was constructed from an immunized alpaca, and one clone recognizing fenitrothion (namely, VHHjd8) was achieved after careful biopanning. It was biotinylated by fusing with the Avi tag and biotin ligase to obtain a fusion protein (VHHjd8-BT), showing both binding capacity to fenitrothion and the streptavidin poly-horseradish peroxidase conjugate (SA-polyHRP). Based on a competitive assay format, the absorbance spectrum of oxidized 3,3',5,5'-tetramethylbenzidine generated by SA-polyHRP overlapped the emission spectrum of carbon dots, which resulted in quenching of signals due to the inner-filter effect. The developed FIA showed an IC
Identifiants
pubmed: 35333506
doi: 10.1021/acs.jafc.2c00826
pmc: PMC9484545
mid: NIHMS1836503
doi:
Substances chimiques
Single-Domain Antibodies
0
Streptavidin
9013-20-1
Fenitrothion
W8M4X3Y7ZY
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
4102-4111Subventions
Organisme : NIEHS NIH HHS
ID : P42 ES004699
Pays : United States
Organisme : NIEHS NIH HHS
ID : R35 ES030443
Pays : United States
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