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

Journal Article · Sat Jan 01 04:00:00 EST 2000 · Journal of Chromatography A

DOI:https://doi.org/10.1016/S0021-9673(00)00572-0· OSTI ID:15020786

Goheen, Steven C; Gibbins, Betty M

Protein losses in ion-exchange and hydrophobic interaction HPLC were examined. The supports were allnon-porous, packed in columns of identical dimensions. 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 fibrinogen. 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 were also investigated. We found a linear relationship between the amount of protein recovered and the log of the molecular mass. Retention times also generally increased with temperature for both HIC and IEC. Other trends in retention behavior and recoveries are discussed.

• Research Organization: Pacific Northwest National Laboratory (PNNL), Richland, WA (US)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 15020786
• Report Number(s): PNNL-SA-45170
• Journal Information: Journal of Chromatography A, Journal Name: Journal of Chromatography A Vol. 890
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ALBUMINS
ANIONS
CATTLE
CHROMATOGRAPHY
CYTOCHROMES
DIMENSIONS
FIBRINOGEN
HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY
Hydrophobic interaction chromatography
IMMUNOGLOBULINS
ION EXCHANGE
PROTEINS
RETENTION
TEMPERATURE DEPENDENCE
proteins