Development of a recombinant Fab-fragment based electrochemical immunosensor for deoxynivalenol detection in food samples.
A reliable and cost-effective electrochemical method for the detection of deoxynivalenol (DON) in cereals and cereal-based food samples based on the use of a novel anti-DON Fab fragment is presented. The analytical system employed, Enzyme-Linked-Immunomagnetic-Electrochemical (ELIME) assay, is based on the use of immunomagnetic beads (IMBs) coupled with eight magnetized screen-printed electrodes (8-mScPEs) as electrochemical transducers. Using standard solutions of DON, a working range between 100 and 4500 ng/ml was obtained with an EC(50) of 380 ng/ml. The ELIME assay was employed to evaluate the cross-reactivity of the Fab fragment towards different trichothecenes revealing a good selectivity towards DON over other trichothecenes with the exception of 3-Ac-DON. The sensor was then applied to cereals and cereal-based food samples (wheat, breakfast cereal and baby-food) and a wide range of sample treatment procedures was tested. Within-laboratory precision (9-24% repeatability for breakfast cereals and 10-33% for baby-food) and recovery data (82-110% for breakfast cereals and 97-108% for baby-food) were calculated by analyzing blank breakfast cereals and baby-foods fortified with DON, demonstrating that the proposed method has the capability for use as a screening assay for DON in such products.
Romanazzo
Daniela
D
Università di Roma, Tor Vergata, Dipartimento di Scienze e Tecnologie Chimiche, Rome, Italy.
Ricci
Francesco
F
Volpe
Giulia
G
Elliott
Christopher T
CT
Vesco
Silvia
S
Kroeger
Katy
K
Moscone
Danila
D
Stroka
Joerg
J
Van Egmond
Hans
H
Vehniäinen
Markus
M
Palleschi
Giuseppe
G
D000818:Animals; D055910:Antibodies, Immobilized; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D002523:Edible Grain / Q000737:chemistry; D055664:Electrochemical Techniques; D005506:Food Contamination / Q000032:analysis*; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007140:Immunoglobulin Fab Fragments; D007223:Infant; D007225:Infant Food / Q000032:analysis; D011994:Recombinant Proteins; D014255:Trichothecenes / Q000032:analysis* / Q000276:immunology; D014908:Triticum / Q000737:chemistry
D055910:Antibodies, Immobilized; D007140:Immunoglobulin Fab Fragments; D011994:Recombinant Proteins; D014255:Trichothecenes; C007262:deoxynivalenol