In vitro synthesis of a type A trichothecenes complete antigen from T-2 toxin

Abstract

We report the artificial synthesis from T-2 toxin of a type A trichothecenes complete antigen. First, 3-Ac-T-2 was made from T-2 following acetylation. Then 3-Ac-NEOS as a T-2 skeleton structure was synthesized by enzymolysis and identified by liquid chromatography-tandem mass spectrometry and nuclear magnetic resonance. Next, a hapten (3-Ac-NEOS-HS) was formed by modifying the C8 position of 3-Ac-NEOS using succinic anhydride method. 3-Ac-NEOS-HS-BSA/-OVA were prepared by conjugating hapten with bovine serum albumin (BSA) or ovalbumin (OVA) by carbodiimide method and identified by UV spectroscopy and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Results showed that conjugation ratio of 3-Ac-NEOS-HS to BSA was 8.76: 1 and OVA 7.24: 1, indicating 3-Ac-NEOS-HS-BSA as a complete antigen was better. Next we used it to immunize rabbits and obtained a 1:64,000 antibody titre. In conclusion, a type A trichothecenes complete antigen was successfully synthesized, which was the foundation for antibody preparation and enzyme-linked immunosorbent assay kit development for all type A trichothecenes (parent + modified/masked type A trichothecenes).