Protein losses in ion-exchange and hydrophobic interaction high-performance liquid chromatography

Protein losses in ion-exchange and hydrophobic interaction HPLC were examin ed. The supports were all non-porous, packed in columns of identical dimens ions. Two ion-exchange chromatography (IEC), anion and cation, as well as a hydrophobic interaction chromatography (HIC) columns were tested. Proteins included cytochrome c, bovine serum albumin (BSA), immunoglobulin G and fi brinogen. Temperature effects on HIC supports were studied for cytochrome c and BSA. Both retention times and recoveries of the proteins were measured . The influence of column residence time on the recovery of proteins was al so investigated. We found a linear relationship between the amount of prote in recovered and the log of the molecular mass. Retention times also genera lly increased with temperature for both HIC and IEC. Other trends in retent ion behavior and recoveries are discussed. (C) 2000 Elsevier Science B.V. A ll rights reserved.