2019
DOI: 10.2144/btn-2018-0143
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Aflatoxin-specific Monoclonal Antibody Selection for Immunoaffinity Column Development

Abstract: Antibodies are the basic components of immunoanalytical systems used for detection of a wide range of analytes. Although there are some ground rules for antibody selection, analyte- and assay-specific criteria are the ones that determine the ultimate success of the immunoassays. In this study, we introduced an effective antibody selection procedure for the development of immunoaffinity columns for aflatoxins. The designed scheme puts emphasis on solvent- and matrix-related characterization steps and was used t… Show more

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Cited by 7 publications
(6 citation statements)
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“…The development of antibody-based immunoaffinity columns comprises the production of the antibodies, immobilization of the antibodies on inert supporting beads such as agarose gel or silica and packing of the beads in cartridges filled with phosphate buffer to maintain physicochemical properties of the antibodies. The evaluation of immunoaffinity columns focus on, but not are limited to, aflatoxin binding capacity and specificity, shelf life, solvent tolerance, cross-reactivity with matrix interferences and reversibility [ 123 , 124 , 125 ]. For sample clean-up, the initial sample extract is loaded on the immunoaffinity column.…”
Section: Sample Preparationmentioning
confidence: 99%
“…The development of antibody-based immunoaffinity columns comprises the production of the antibodies, immobilization of the antibodies on inert supporting beads such as agarose gel or silica and packing of the beads in cartridges filled with phosphate buffer to maintain physicochemical properties of the antibodies. The evaluation of immunoaffinity columns focus on, but not are limited to, aflatoxin binding capacity and specificity, shelf life, solvent tolerance, cross-reactivity with matrix interferences and reversibility [ 123 , 124 , 125 ]. For sample clean-up, the initial sample extract is loaded on the immunoaffinity column.…”
Section: Sample Preparationmentioning
confidence: 99%
“…We used the cross-reactions test (CR) as described by Ertekin et al [36]. According to the CR, ZEN and its structural analogs, including α-ZAL, β-ZAL, α-ZOL, β-ZOL, and ZON, were selected as inhibitors.…”
Section: Specificity Measurementmentioning
confidence: 99%
“…The CR value of the ZEN mAbs, indicating their respective specificities, was determined using an icELISA for ZEN and its five homologs (α-ZAL, β-ZAL, α-ZEL, β-ZEL, and ZAN), and other common mycotoxins, such as AFB1, DON, FB1, OTA, and T-2 toxin. The CR value was calculated as follows: CR (%) = [IC50 (ZEN)/IC50 (competitor)] × 100% [ 40 , 41 ]. The ZEN mAb with the highest titer, the maximum affinity, the lowest IC50 value, and the smallest CR value was filtrated for the development of the icELISA.…”
Section: Methodsmentioning
confidence: 99%