Aflatoxin-specific monoclonal antibody selection for immunoaffinity column development

Antibodies are the basic components of immuno-analytical 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 to comparatively evaluate eight monoclonal antibodies. The selected antibody was tolerant to 40% methanol, 20% acetonitrile, 30% acetone and 40% ethanol and did not interact with corn, red pepper or hazelnut extracts. Immunoaffinity columns developed with the selected antibody were validated by 15 independent aflatoxin analysis laboratories. METHOD SUMMARY This article describes a systematic methodology to select the best monoclonal antibody to be used in aflatoxin immunoaffinity columns. The scheme consists of four critical steps. Highly specific antibodies were selected that interact with aflatoxin derivatives but not with other mycotoxins. High tolerance to the solvents used in aflatoxin extraction protocols was assured. The antibodies that interact with food extracts were eliminated. Finally, the antibodies were functionally tested for effective integration to immunoaffinity columns where resin binding efficacy and column performances were evaluated. The developed columns were validated by 15 independent laboratories to show the efficacy of the selection scheme.