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.
Ohtani
K
K
Department of Toxicology and Microbial Chemistry, Faculty of Pharmaceutical Sciences, Science University of Tokyo, Japan.
Kawamura
O
O
Ueno
Y
Y
D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D006863:Hydrogen-Ion Concentration; D051379:Mice; D011506:Proteins / Q000032:analysis / Q000302:isolation & purification*; D011863:Radioimmunoassay; D012717:Sesquiterpenes / Q000302:isolation & purification*; D013605:T-2 Toxin / Q000032:analysis / Q000302:isolation & purification*; D013696:Temperature
D011506:Proteins; D012717:Sesquiterpenes; D013605:T-2 Toxin