Improved preparation of T-2 toxin-protein conjugates

T-2 toxin (T-2) was converted into T-2 hemisuccinate (T-2 HS), T-2 hemiglutarate (T-2 HG) and T-2 hemiphthalate (T-2 HP), and conjugated with bovine serum albumin (BSA), ovalbumin (OVA) and keyhole limpet haemocyanin (KLH) using carbodiimide under various conditions. The recovery of T-2 conjugated proteins and the amount of T-2 bound were largely dependent on the amount of coupling reagent, pH, reaction temperature, buffer and reaction time. In regard to T-2 derivatives binding with a protein, T-2 HG was the most efficient, followed by T-2 HS and then T-2 HP. With regard to T-2 to protein binding, KLH was the most efficient, followed by BSA and then OVA. To promote T-2-protein binding, phosphate-buffered saline was more effective than cacodylate buffer. Within a pH range of 5.5-7.0, weakly acidic conditions were more efficient than neutral conditions for promoting T-2-protein binding.