Development of a sensitive monoclonal-based enzyme-linked immunosorbent assay for monitoring T-2 toxin in food and feed

ABSTRACT

The consumption of food or feed contaminated with high levels of T-2 toxin may cause adverse health effects in humans and other animals. In this study, to monitor T-2 toxin rapidly in food and feed, a sensitive and specific monoclonal antibody (mAb) against T-2 toxin was generated and a simple and rapid indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) developed. T-2 toxin was first converted to T-2-hemisuccinate (T-2HS) and T-2-hemiglutarate (T-2HG), which were then conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to prepare an immunogen and coating antigen, respectively. After the inoculation of female Balb/c mice and cell fusions, one cell line, 4D8, with the IgG1 isotype was obtained. The 4D8 antibody exhibited the ability specifically to recognise T-2 toxin with IC₅₀ 1.46 µg l⁻¹. Based on this 4D8 mAb, an optimised ic-ELISA protocol was developed using only methanol–water (7:3, v/v) in feed and cereal samples and ethyl acetate in muscle samples. The limits of detection of T-2 toxin in various sample matrices varied from 0.07 to 15.8 µg kg⁻¹; the recoveries ranged from 50.3% to 113.6%; and the CVs were less than 19.0%. These results suggest that the prepared mAb and the developed ic-ELISA method will be a useful tool for detecting T-2 toxin in foods and feeds.

KEYWORDS:

• T-2 toxin
• monoclonal antibody
• indirect competitive enzyme-linked immunosorbent assay
• food
• feed

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

The work was financially supported by the Ministry of Science and Technology of the People’s Republic of China [grant number 2011-G5, GJFP2016008] and the Fundamental Research Funds for the Central Universities [grant numbers 2662015QC030 and 2662015PY021].