Determination of fibrinogen in plasma by high-performance immunoaffinity chromatography

Abstract A high-performance immunoaffinity chromatographic (HPIAC) method for fibrinogen was developed which had several advantages over existing methodologies including increased linear range and no interference from heparin. Several modifications of usual HPIAC procedures were necessary including the employment of a methacrylate polymeric support to reduce non-specific adsorption and the addition of urea to a pH 2.1 elution buffer to affect elution. A significant split-peak effect ( i.e. , unretained fibrinogen) was noted at higher flow-rates and at higher fibrinogen concentrations, which was shown to be temperature-dependent, with the amount of fibrinogen retained on the column increasing with increased temperature.