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Generic anti-sulfonamide immunoaffinity columns made using sulfamethazine-specific polyclonal antibodies

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Reusable (>100 times) immunoaffinity columns (IACs) with high capacity (>50 µg sulfonamide/ml gel) were made with affinity purified anti-sulfonamide polyclonal antibodies (pAbs). The specificity and capacity of these IACs for sulfonamide N 1 -analogues and N 4 -derivatives were investigated in competitive and non-competitive environments. Even though antibody affinity toward various sulfonamides differed by as much as 500-fold when measured by competitive enzyme-linked immunosorbent assay (ELISA), we demonstrated that a generic immunoaffinity application was still possible as long as the saturation limit of the column was not exceeded. Anti-sulfamethazine pAbs could equally recover a mixture of sulfonamide compounds under non-competitive conditions by immunoaffinity chromatography; however, under saturating and competitive conditions, IACs could only recover sulfonamides closely resembling the immunogen. This study also presents evidence that ELISA can serve to screen pAbs for use in generic immunoaffinity chromatography of sulfonamides.
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Keywords: Polyclonal antibody; cross-reactivity; generic specificity; immunoaffinity chromatography; linking arm effect; sulfonamide

Document Type: Research Article

Affiliations: Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta, Canada

Publication date: December 1, 2005

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