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Eu3+‐labeled IgG‐based time‐resolved fluoroimmunoassay for highly sensitive detection of aflatoxin B1 in feed

Author:
Hu, Xiaofei, Yao, Jingjing, Wang, Fangyu, Yin, Mengqi, Sun, Yaning, Hu, Mei, Shi, Qiaoqiao, Zhang, Gaiping
Source:
Journal of the science of food and agriculture 2018 v.98 no.2 pp. 674-680
ISSN:
0022-5142
Subject:
aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin M1, cell fusion, cross reaction, detection limit, employment, enzyme-linked immunosorbent assay, feeds, fluorescent antibody technique, high performance liquid chromatography, immunization, immunoglobulin G, inhibitory concentration 50, monoclonal antibodies, toxicity
Abstract:
BACKGROUND: Aflatoxin B₁ (AFB₁) is a kind of toxic and carcinogenic mycotoxin. A time‐resolved fluoroimmunoassay (TRFIA) was established for quantitative detection of AFB₁ in feed using Eu³⁺‐labeled IgG as tracer. RESULTS: Monoclonal antibody (McAb) against AFB₁ (9B11‐D7) was prepared through immunization and cell fusion and was identified as high affinity, specificity and sensibility by enzyme‐linked immunosorbent assay (ELISA). The 50% inhibition value (IC₅₀) was 0.81 ng mL⁻¹, the limit of detection (LOD) was 0.10 ng mL⁻¹ and detection range was 0.10–3.94 ng mL⁻¹. Goat anti‐mouse immunoglobulin G (IgG) was modified by Eu³⁺‐DATT, generating Eu³⁺‐labeled IgG. Under optimal assay conditions, TRFIA was shown to be highly sensitive and specific in detection of AFB₁. The IC₅₀ and LOD were 94.73 pg mL⁻¹ and 3.55 pg mL⁻¹, respectively, and detection range was 3.55–1.11 × 10³ pg mL⁻¹. Cross‐reactivity with AFM₁, AFB₂, AFG₁ and AFG₂ was 31.26%, 37.6%, 127.46% and 35.74%, respectively, but zero with other analogues. In determination of AFB₁ spiked in feed sample, TRFIA showed high accuracy and precision. The average recoveries ranged from 93.71% to 97.80%, and coefficient of variation was 1.25–3.73%. Good correlation between TRFIA and HPLC was demonstrated for determination of AFB₁ in feeds, confirming the reliability of the developed method. CONCLUSION: The developed TRFIA exhibited good potential for employment in the ultrasensitive detection of AFB₁ in feed and could be used to determine total aflatoxins. © 2017 Society of Chemical Industry
Agid:
5872135