Fractionation of plasma proteins using dye-ligand chromatography: elution profiles on immobilized green TSK-AF.
The fractionation of human plasma by chromatography on immobilized Green TSK-AF was assessed by immunological analysis of the elution profiles of 27 different plasma proteins. A three-step procedure was used to elute proteins from the column. First a low-molarity buffer (30 mM sodium phosphate, pH 7.0, I = 0.053) was applied; then a linear salt gradient (0-1.0 M NaCl in the above buffer) was followed by an additional wash with four bed volumes of 1.0 M NaCl. Tightly bound proteins were finally stripped with 0.5 M NH4SCN. The elution profile of the proteins using this procedure appears to be very reproducible. Comparison with the profile obtained upon chromatography on Cibacron Blue 3GA [Gianazza, E. and Arnaud, P. (1982) Biochem. J. 201, 129-136] indicates significant differences between the binding properties of the two gels. These differences can be used to design a "tandem-chromatography" system which provides an efficient means for the separation of several plasma proteins.
Department of Microbiology and Immunology, Medical University of South Carolina, Charleston 29425.
L
L
Miribel
P
P
Arnaud
D001798:Blood Proteins / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D004396:Coloring Agents; D006801:Humans; D007122:Immunoelectrophoresis / Q000379:methods; D007202:Indicators and Reagents; D013056:Spectrophotometry, Ultraviolet
D001798:Blood Proteins; D004396:Coloring Agents; D007202:Indicators and Reagents