Rapid size-exclusion chromatography of proteoglycans

The separation of intact proteoglycans using high-performance liquid chromatography is not trivial because the high molarity denaturing buffers required to maintain proteoglycans in the disaggregated state create back-pressures higher than the limits of many HPLC systems. Until recently, low back-pressure requirements of HPLC size-exclusion columns precluded their use for the separation of intact proteoglycans. In this study we show that rapid size-exclusion chromatography is possible in 8 M urea buffers using a Dionex BioLC system equipped with a Bio-Rad BioSil Sec-400 column. This technique reduced the time required for size-exclusion chromatography of intact proteoglycans from approximately 18 h (Sepharose CL4B) to 25 min and in some cases improved resolution of the sample.