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Preparative size-exclusion chromatography of human serum apolipoproteins using an analytical liquid chromatograph
Authors:CTimothy Wehr  Robert L Cunico  Gary S Ott  Virgie G Shore
Institution:1. Varian Instrument Group, Walnut Creek Instrument Division, 2700 Mitchell Drive, Walnut Creek, California 94598 USA;2. Lawrence Livermore National Laboratory, Biomedical Sciences Division, University of California, Livermore, California 94550 USA
Abstract:Apolipoproteins, extracted from human serum high-density lipoproteins, can be resolved and recovered with high yield from a preparative MicroPak TSK Type 3000SW size-exclusion column using Tris-buffered 6 m urea or 6 m guanidinium chloride mobile phases. Adequate resolution of some apolipoprotein pairs is only achieved at low flow velocities and low sample loads, necessitating repetitive injections of small amounts of material for preparative isolation. An analytical high-performance liquid chromatograph equipped with a simplified sample introduction scheme and low-pressure switching valves for fraction collection was used to isolate milligram quantities of HDL apolipoproteins.
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