High-performance liquid chromatography of proteins on compressed, non-porous agarose beads. I. Hydrophobic-interaction chromatography.
Macroporous agarose beads were converted into non-porous beads by shrinkage and cross-linking in organic solvents. These beads could be used for high-performance hydrophobic-interaction chromatography without derivatization with non-polar ligands, because the 1,4-butanediol diglycidyl ether, used as cross-linker, gives relatively hydrophobic bridges. The resolution for compressed columns packed with these beads was determined as a function of gradient time at constant flow-rate, flow-rate at constant gradient volume and flow-rate at constant gradient time and as a function of loading capacity. Interestingly, the resolution is virtually independent of flow-rate at constant gradient volume even when the column is packed with relatively large beads (diameter 30 microns). The beads have the advantage of being stable up to pH 14.
Hjertén
S
S
Institute of Biochemistry, University of Uppsala, Sweden.
Liao
J L
JL
D002072:Butylene Glycols; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents; D011506:Proteins / Q000302:isolation & purification*; D012685:Sepharose
D002072:Butylene Glycols; D003432:Cross-Linking Reagents; D011506:Proteins; D012685:Sepharose; C014376:1,4-bis(2,3-epoxypropoxy)butane