PMID- 36287981
OWN - NLM
STAT- MEDLINE
DCOM- 20221028
LR  - 20221222
IS  - 2072-6651 (Electronic)
IS  - 2072-6651 (Linking)
VI  - 14
IP  - 10
DP  - 2022 Oct 19
TI  - Nanobody-Nanoluciferase Fusion Protein-Enabled Immunoassay for Ochratoxin A in 
      Coffee with Enhanced Specificity and Sensitivity.
LID - 10.3390/toxins14100713 [doi]
LID - 713
AB  - Ochratoxin A (OTA), one of the best-known mycotoxins, causes problems concerning 
      food safety with potential toxic effects in humans and animals. So, it is crucial 
      to develop simple and sensitive methods for the detection of OTA. Herein, a 
      nanoluciferase-nanobody fusion protein (Nb28-Nluc)-retaining antibody recognition 
      and enzymatic activity was first prepared, which was then applied as a 
      bifunctional tracer to construct a one-step bioluminescent enzyme-linked 
      immunosorbent assay (BLEIA) for OTA in coffee samples. On the basis of Nb28-Nluc, 
      the BLEIA can be completed with a one-step incubation and detection, with only a 
      substrate replacement from 3,3',5,5'-tetramethylbenzidine (TMB) to a Nluc assay 
      reagent (Furimazine). Under the optimal experimental conditions, the proposed 
      one-step BLEIA achieved a detection limit of 3.7 ng/mL (IC(10)) within 3 h. 
      Moreover, the BLEIA method showed good repeatability and accuracy in the spike 
      recovery experiments with recoveries of 83.88% to 120.23% and relative standard 
      deviations (RSDs) of 5.2% to 24.7%, respectively. Particularly, the BLEIA 
      displayed superior performances, such as fewer operations and more rapid and 
      sensitive detection as compared with Nb28-based enzyme-linked immunosorbent 
      assay. Therefore, the proposed one-step BLEIA has great potential for the 
      sensitive and accurate screening of OTA in food samples.
FAU - Bao, Kunlu
AU  - Bao K
AD  - Key Laboratory of Tropical and Vegetables Quality and Safety for State Market 
      Regulation, School of Food Science and Engineering, Hainan University, Haikou 
      570228, China.
FAU - Liu, Xing
AU  - Liu X
AUID- ORCID: 0000-0002-1301-4837
AD  - Key Laboratory of Tropical and Vegetables Quality and Safety for State Market 
      Regulation, School of Food Science and Engineering, Hainan University, Haikou 
      570228, China.
FAU - Liao, Yujing
AU  - Liao Y
AD  - Key Laboratory of Tropical and Vegetables Quality and Safety for State Market 
      Regulation, School of Food Science and Engineering, Hainan University, Haikou 
      570228, China.
FAU - Liu, Zilong
AU  - Liu Z
AD  - Key Laboratory of Tropical and Vegetables Quality and Safety for State Market 
      Regulation, School of Food Science and Engineering, Hainan University, Haikou 
      570228, China.
FAU - Cao, Hongmei
AU  - Cao H
AD  - Key Laboratory of Tropical and Vegetables Quality and Safety for State Market 
      Regulation, School of Food Science and Engineering, Hainan University, Haikou 
      570228, China.
FAU - Wu, Long
AU  - Wu L
AUID- ORCID: 0000-0003-0212-6419
AD  - Key Laboratory of Tropical and Vegetables Quality and Safety for State Market 
      Regulation, School of Food Science and Engineering, Hainan University, Haikou 
      570228, China.
FAU - Chen, Qi
AU  - Chen Q
AD  - National Engineering Research Center for Bioengineering Drugs and the 
      Technologies, Institute of Translational Medicine, Nanchang University, Nanchang 
      330031, China.
AD  - Key Laboratory of Tropical and Vegetables Quality and Safety for State Market 
      Regulation, School of Food Science and Engineering, Hainan University, Haikou 
      570228, China.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20221019
PL  - Switzerland
TA  - Toxins (Basel)
JT  - Toxins
JID - 101530765
RN  - 1779SX6LUY (ochratoxin A)
RN  - 0 (Coffee)
RN  - 0 (Ochratoxins)
RN  - 0 (Mycotoxins)
SB  - IM
MH  - Humans
MH  - Animals
MH  - Coffee
MH  - *Ochratoxins/analysis
MH  - Immunoassay
MH  - *Mycotoxins/analysis
MH  - Sensitivity and Specificity
MH  - Food Contamination/analysis
PMC - PMC9609117
OTO - NOTNLM
OT  - BLEIA
OT  - Nb28-Nluc
OT  - bioluminescent
OT  - food safety
OT  - ochratoxin A
COIS- The authors declare no conflict of interest.
EDAT- 2022/10/27 06:00
MHDA- 2022/10/29 06:00
PMCR- 2022/10/19
CRDT- 2022/10/26 12:43
PHST- 2022/09/20 00:00 [received]
PHST- 2022/10/05 00:00 [revised]
PHST- 2022/10/18 00:00 [accepted]
PHST- 2022/10/26 12:43 [entrez]
PHST- 2022/10/27 06:00 [pubmed]
PHST- 2022/10/29 06:00 [medline]
PHST- 2022/10/19 00:00 [pmc-release]
AID - toxins14100713 [pii]
AID - toxins-14-00713 [pii]
AID - 10.3390/toxins14100713 [doi]
PST - epublish
SO  - Toxins (Basel). 2022 Oct 19;14(10):713. doi: 10.3390/toxins14100713.