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High affinity antibody based on a rationally designed hapten and development of a chemiluminescence enzyme immunoassay for quantification of Alternariol in fruit Juice, maize and flour

Yao, Chan-Yuan, Xu, Zhen-Lin, Wang, Hong, Zhu, Fan, Luo, Lin, Yang, Jin-Yi, Sun, Yuan-Ming, Lei, Hong-Tao, Tian, Yuan-Xin, Shen, Yu-Dong
Food chemistry 2019 v.283 pp. 359-366
Alternaria, antibodies, chemiluminescence, corn, cross reaction, detection limit, enzyme immunoassays, flour, fruit juices, haptens, high performance liquid chromatography, mutagens, mycotoxins, screening, secondary metabolites, tandem mass spectrometry
Alternariol (AOH), a secondary metabolites of the genus Alternaria, is a genotoxic mycotoxin. Because the previous “Mannich antigen” for AOH readily underwent unstable induction, a new AOH hapten was designed for inducing a high-quality antibody against AOH using a conjugate of the AOH carboxymethyl derivative with a carrier protein as the immunogen. A competitive indirect chemiluminescence enzyme immunoassay (ciCLEIA) for AOH was optimized and showed good sensitivity (limit of detection 0.068 ng/mL), a linear range (0.11–1.23 ng/mL), and negligible cross-reactivity with analogues. Average rates of recovery from spiked samples (juice and cereal) were between 72.7% and 115.8%, with coefficients of variation <14.3%. Results from ciCLEIA correlated well with those of high-performance liquid chromatography coupled with tandem mass spectrometry, meaning the proposed method might be an effective screening tool for detection of AOH in food samples.