Nan.obody-based enzyme immunoassay for ochratoxin A in cereal with high resistance to matrix interference

被引：74

作者：

Liu, Xing ^{[1
,2
]}

Tang, Zongwen ^{[1
]}

Duan, Zhenhua ^{[2
]}

He, Zhenyun ^{[3
]}

Shu, Mei ^{[4
]}

Wang, Xianxian ^{[4
]}

Gee, Shirley J. ^{[5
,6
]}

Hammock, Bruce D. ^{[5
,6
]}

Xu, Yang ^{[4
]}

机构：

[1] Hainan Univ, Coll Food Sci & Technol, 58 Renmin Ave, Haikou 570228, Peoples R China

[2] Hezhou Univ, Inst Food Res, Hezhou 542899, Peoples R China

[3] Natl Univ Singapore, Dept Chem, Food Sci & Technol Program, 3 Sci Dr 3, Singapore 117543, Singapore

[4] Nanchang Univ, State Key Lab Food Sci & Technol, 235 Nanjing East Rd, Nanchang 330047, Jiangxi, Peoples R China

[5] Univ Calif Davis, Dept Entomol & Nematol, Davis, CA 95616 USA

[6] Univ Calif Davis, Ctr Comprehens Canc, Davis, CA 95616 USA

来源：

TALANTA | 2017年 / 164卷

关键词：

Nanobody; Ochratoxin A; Immunoassay; Cereal; Matrix interference; FUSION PROTEIN; FUMONISIN B-1; FV-FRAGMENTS; ANTIBODIES; VHH; NANOBODY; EXPRESSION; AFLATOXIN; AFFINITY; ALPACA;

D O I：

10.1016/j.talanta.2016.11.039

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A sensitive indirect competitive nanobody-based enzyme linked immunosorbent assay (Nb-ELISA) for ochratoxin A (OTA) with high resistance to cereal matrix interference was developed. Nanobodies against OTA (Nb15, Nb28, Nb32, Nb36) were expressed in E. coil cells and their thermal stabilities were compared with that of an OTA-specific monoclonal antibody 6H8. All nanobodies could still retain their antigen-binding activity after exposure to temperature 95 degrees C for 5 min or to 90 degrees C for 75 min. Nb28 that exhibited the highest sensitivity in ELISA was selected for further research. An indirect competitive ELISA based on Nb28 was developed for OTA, with an IC50 of 0.64 ng/mL and a linear range (IC20-IC80) of 0.27-1.47 ng/mL. Cereal samples were analyzed following a 2.5 fold dilution of sample extracts, showing the good resistance to matrix interference of the Nb-ELSLk. The recovery of spiked cereal samples (rice, oats, barley) ranged from 80% to 105% and the Nb-ELISA results of OTA content in naturally contamined samples were in good agreement with those determined by a commercial ELISA kit. The results indicated the reliablity of nanobody as a promising immunoassay reagent for detection of mycotoxins in food matrix and its potential in biosensor development.

引用

页码：154 / 158

页数：5

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