Monoclonal antibody production and the development of an indirect competitive enzyme-linked immunosorbent assay for screening T-2 toxin in milk. (15th December 2018)

• Record Type: Journal Article
• Authors: Xu, Huijing; Dong, Yuan; Guo, Jian; Jiang, Xiaoming; Liu, Jing; Xu, Song; Wang, Huiyan
• Is Part Of: Toxicon. Volume 156(2018)
• Page Start: 1
• Page End: 6
• Publication Date: 2018-12-15
• Subjects: T-2 toxin -- Highly specific monoclonal antibody -- ELISA -- Milk; Toxins -- Periodicals; Venom -- Periodicals; 615.9
• DOI: 10.1016/j.toxicon.2018.10.307
• Languages: English
• ISSNs: 0041-0101

Abstract

Abstract: To monitor T-2 toxin rapidly in milk, a highly specific against T-2 toxin monoclonal antibody (mAb) was produced and a sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed. T-2 toxin was first converted to T-2-adipic anhydride (T-2HA), and then Keyhole limpet hemocyanin (KLH) and virus-like particles (VLP) were synthesized and conjugated to T-2HA as coating and immunogen antigen, respectively. After the inoculation of female Balb/c mice and cell fusions, a high affinity mAb was produced. After optimization, the 50% inhibition concentration (IC50 ) for the developed ic-ELISA was estimated as 0.137 ng mL −1 . Based on this mAb, an optimized ic-ELISA protocol was developed using the dilution method to prepare milk samples. The limits of detection of T-2 toxin in milk was 0.827 ng mL −1, the mean recovery of T-2 toxin in spiked samples (2.5, 5 and 10 ng mL −1 ) ranged from 85.0% to 98.9%, and the CVs ranged from 6.1% to 8.6%. The ic-ELISA was validated by HPLC-MS/MS method, and all results demonstrated that it was a suitable screening method for detecting T-2 toxin in milk. Highlights: A highly specific against T-2 toxin monoclonal antibody (mAb) is produced. A sensitive ic-ELISA method to screen T-2 toxin in milk is developed. The limit of detection of T-2 toxin in milk is determined. The ic-ELISA is validated by HPLC-MS/MS method.