Procedure for radiolabeling gizzerosine and basis for a radioimmunoassay
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Date
1999
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Abstract
A method for the labeling of gizzerosine (GZ), a biogenic amine found in fish meal, is described. The labeling procedure with I-125 using a water-soluble Bolton-Hunter reagent and a mild water-insoluble oxidant (Iodogen) reagent is rapid and reproducible. The I-125-GZ hapten was demonstrated to be immunologically active in a radioimmunoassay developed with polyclonal antibodies to GZ absorbed with a histamine-Sepharose column. The curves were linear in the range of 0.0001 and 0.1 mu g/mL. Samples of fish meal previously extracted of histamine with methanol and submitted to acid hydrolysis were contaminated with known amounts of GZ and submitted to the assay. The fish meal samples contaminated with GZ show a dose-response effect similar to the standard curve, and apparently the other component present in the sample did not interfere with the binding of the antibodies to I-125-GZ. These data indicate the suitability of the radioimmunoassay to determine specifically GZ in fish meal.
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Keywords
gizzerosine labeling, polyclonal antibodies, RIA