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

Author: Hao Zhang 1, 2, Hai-zhen Mo 2* and Hua Li 3*
Received 29 September 2012, accepted 20 January 2013.
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 AFM 2 (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.

Journal: Food, Agriculture and Environment (JFAE)
Online ISSN: 1459-0263Year: 2013, Vol. 11, Issue 1, pages 190-194. Publisher: WFL.


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