RAPID SIZE-EXCLUSION CHROMATOGRAPHY OF PROTEOGLYCANS

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