Determination of alkaline phosphatase aggregation by size exclusion high-performance liquid chromatography with low-angle laser light scattering detection

Abstract The formation of aggregates of alkaline phosphatase (AP) is examined in two different buffer systems: phosphate-buffered saline and modified protein buffer, using isocratic size-exclusion chromatography combined with low-angle laser-light scattering, ultraviolet spectroscopy, and a modified differential refractive index detector, all in series. The effects of buffer conditions, concentrations, and different batch preparations of alkaline phosphatase, as obtained from a manufacturer, were examined.