Abstract
A facile method for activating hydroxyl groups of Sepharose gel using 2-fluoro-1-methylpyridinium toluene-4-sulfonate (FMP) in the presence of triethylamine is described. Nucleophilic affinity ligands, i.e., primary amino or sulfhydryl bearing compounds in slightly alkaline (pH 8-9) aqueous solutions or in organic solvents can be coupled directly to the activated gel. Such nucleophilic reactions result in displacing the pyridinium moiety from the activated gel as 1-methyl-2-pyridone and in forming stable, non-ionic secondary amine or thioether linkages between the gel and the ligands being immobilized. Because 1-methyl-2-pyridone has a molar extinction coefficient of 5,900 at 297 nm, the activation and coupling reactions can be followed conveniently by a simple spectrophotometric method.
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Ngo, T. Facile Activation of Sepharose Hydroxyl Groups by 2-Fluoro-1 Methylpyridinium Toluene-4-Sulfonate: Preparation of Affinity and Covalent Chromatographic Matrices. Nat Biotechnol 4, 134–137 (1986). https://doi.org/10.1038/nbt0286-134
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DOI: https://doi.org/10.1038/nbt0286-134
