Journal of Food, Agriculture and Environment

Preparation of aflatoxin B₁ polyclonal antibody and development of an indirect competitive enzyme-linked immunosorbent assay

Author(s):

Hao Zhang ^{1, 2}, Hai-zhen Mo ²*, Hua Li ³*

Recieved Date: 2012-09-29, Accepted Date: 2013-01-20

Abstract:

This article aimed to prepare polyclonal antibodies (pAbs) against aflatoxin B₁ (AFB₁) and develop an immunoassay for detecting aflatoxin residues in tea matrix. After derivation, AFB₁ hapten was conjugated to carrier proteins to synthesize the artificial antigen, and rabbits were employed to produce pAbs. Based on the bidimensional checkerboard titration results, two indirect competitive ELISA (icELISA) standard curves have been established. For the traditional two-step assay, the linear range was from 0.12 to 103 ng/ml, with the half maximal inhibitory concentration (IC₅₀) and limit of detection (LOD) values of 2.97 and 0.05 ng/ml, respectively; while the rapid one-step icELISA had a working range from 0.14 to 126 ng/ml, with IC₅₀ and LOD values of 3.85 and 0.06 ng/ml. Of all the competitive analogues, this one-step assay exhibited a high cross-reactivity to AFB₂ (26.2%), AFG₁ (61.3%), AFG₂ (25.5%), AFM₁ (18.6%) and AFM₂ (3.4%). It also indicated that a 10-fold dilution in tea extracts gave an inhibition curve almost the same as that in PBS buffer. These results suggest that the immunoassay provided was a promising alternative for screening aflatoxin residues in tea sample, even in other agro-products such as peanuts, feedstuffs and vegetable oils. 

Keywords:

Aflatoxin B₁, artificial antigen, polyclonal antibody, indirect competitive ELISA, tea sample

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Journal: Journal of Food, Agriculture and Environment
Year: 2013
Volume: 11
Issue: 1
Category: Food and Health
Pages: 190-194

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