Peer-Reviewed Journal Details
Mandatory Fields
Edupuganti S.;Edupuganti O.;O'Kennedy R.;Defrancq E.;Boullanger S.
2013
April
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Use of T-2 toxin-immobilized amine-activated beads as an efficient affinity purification matrix for the isolation of specific IgY
Published
7 ()
Optional Fields
Affinity purification Enzyme-linked immunoassay (ELISA) SPR-based inhibition assay T-2 toxin immobilized amine-activated beads
923-924
98
101
An affinity purification method that isolates T-2 toxin-specific IgY utilizing a T-2-toxin-immobilized column was developed. The T-2 toxin was covalently coupled via a carbonyldiimidazole-activated hydroxyl functional group to amine-activated sepharose beads. The affinity-purified IgY was characterized by gel electrophoresis, fast protein liquid chromatography, enzyme-linked immunosorbant assay, surface plasmon resonance and mass spectrometry. A competitive inhibition ELISA (CI-ELISA) was performed using affinity-purified IgY with a T-2 toxin detection sensitivity of 30ng/mL, which falls within the maximum permissible limit of 100ng/mL. The cross reactivity of IgY towards deoxynivalenol, zearalenone, fumonisin B1 and HT-2 was significantly reduced after affinity purification. A surface plasmon resonance (SPR)-based inhibition assay was also applied for quantitative determination of T-2 toxin in spiked wheat samples. The results obtained indicate the feasibility of utilizing this IgY-based assay for the detection of T-2 toxin in food samples. © 2013 .
1570-0232
10.1016/j.jchromb.2013.02.007
Grant Details