Use of T-2 toxin-immobilized amine-activated beads as an efficient affinity purification matrix for the isolation of specific IgY
| Autor: | Stéphanie Boullanger, Om Prakash Edupuganti, Richard O'Kennedy, Eric Defrancq, Soujanya Ratna Edupuganti |
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| Přispěvatelé: | inconnu, Inconnu, Centre de Recherches sur les Macromolécules Végétales (CERMAV), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), Carret, Michèle |
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Clinical Biochemistry
Immunoglobulins Enzyme-Linked Immunosorbent Assay medicine.disease_cause 01 natural sciences Biochemistry Sensitivity and Specificity Chromatography Affinity Mass Spectrometry Analytical Chemistry Sepharose Matrix (chemical analysis) 03 medical and health sciences Non-competitive inhibition Affinity chromatography medicine Animals Surface plasmon resonance Triticum ComputingMilieux_MISCELLANEOUS Gel electrophoresis 0303 health sciences Chromatography 030306 microbiology Chemistry Toxin 010401 analytical chemistry Reproducibility of Results Fast protein liquid chromatography Cell Biology General Medicine Surface Plasmon Resonance 0104 chemical sciences T-2 Toxin Electrophoresis Polyacrylamide Gel Chickens |
| Zdroj: | Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, Elsevier, 2013, pp.98-101 Europe PubMed Central |
| ISSN: | 1570-0232 1873-376X |
| Popis: | 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 30 ng/mL, which falls within the maximum permissible limit of 100 ng/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. |
| Databáze: | OpenAIRE |
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