TY - JOUR
T1 - Regenerable immuno-biochip for screening ochratoxin A in green coffee extract using an automated microarray chip reader with chemiluminescence detection
AU - Sauceda-Friebe, Jimena C.
AU - Karsunke, Xaver Y.Z.
AU - Vazac, Susanna
AU - Biselli, Scarlett
AU - Niessner, Reinhard
AU - Knopp, Dietmar
N1 - Funding Information:
We gratefully acknowledge financial support by the Federal Ministry of Education and Research (BMBF, project BIOFOCOS). Further, the authors thank Dr. G. Lystik (Soft Flow Biotechnology, Gödöllö, Hungary) for providing the anti-OTA monoclonal antibody. We are also grateful to Susanna Mahler for IAC clean-up and HPLC measurements, York Karsunke (Softwarebüro Karsunke, Wolnzach, Germany) for the free license of SIP0.4, and Huntsman Corporation for the free samples of DAPEG.
PY - 2011/3/18
Y1 - 2011/3/18
N2 - Ochratoxin A (OTA) can contaminate foodstuffs in the ppb to ppm range and once formed, it is difficult to remove. Because of its toxicity and potential risks to human health, the need exists for rapid, efficient detection methods that comply with legal maximum residual limits. In this work we have synthesized an OTA conjugate functionalized with a water-soluble peptide for covalent immobilization on a glass biochip by means of contact spotting. The chip was used for OTA determination with an indirect competitive immunoassay format with flow-through reagent addition and chemiluminescence detection, carried out with the stand-alone automated Munich Chip Reader 3 (MCR 3) platform. A buffer model and real green coffee extracts were used for this purpose. At the present, covalent conjugate immobilization allowed for at least 20 assay-regeneration cycles of the biochip surface. The total analysis time for a single sample, including measurement and surface regeneration, was 12min and the LOQ of OTA in green coffee extract was 0.3μgL-1 which corresponds to 7μgkg-1.
AB - Ochratoxin A (OTA) can contaminate foodstuffs in the ppb to ppm range and once formed, it is difficult to remove. Because of its toxicity and potential risks to human health, the need exists for rapid, efficient detection methods that comply with legal maximum residual limits. In this work we have synthesized an OTA conjugate functionalized with a water-soluble peptide for covalent immobilization on a glass biochip by means of contact spotting. The chip was used for OTA determination with an indirect competitive immunoassay format with flow-through reagent addition and chemiluminescence detection, carried out with the stand-alone automated Munich Chip Reader 3 (MCR 3) platform. A buffer model and real green coffee extracts were used for this purpose. At the present, covalent conjugate immobilization allowed for at least 20 assay-regeneration cycles of the biochip surface. The total analysis time for a single sample, including measurement and surface regeneration, was 12min and the LOQ of OTA in green coffee extract was 0.3μgL-1 which corresponds to 7μgkg-1.
KW - Automated immunoassay
KW - Chemiluminescence detection
KW - Green coffee
KW - Ochratoxin A (OTA)
KW - Peptide-OTA conjugate
KW - Regenerable biochip
UR - http://www.scopus.com/inward/record.url?scp=79952485291&partnerID=8YFLogxK
U2 - 10.1016/j.aca.2011.01.030
DO - 10.1016/j.aca.2011.01.030
M3 - Article
C2 - 21397079
AN - SCOPUS:79952485291
SN - 0003-2670
VL - 689
SP - 234
EP - 242
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
IS - 2
ER -