Fractionation of plasma proteins by pseudo-ligand affinity chromatography. Elution from immobilized Brown Fractogel TSK-AF.
The fractionation of human plasma by chromatography on immobilized Brown Fractogel TSK-AF was analyzed by following the elution profile of 25 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 sodium chloride in the above buffer) was followed by an additional wash with four bed volumes of 1.0 M sodium chloride. Tightly bound proteins were finally stripped with 0.5 M ammonium thiocyanate. The elution profile of the proteins obtained by this procedure appears to be very reproducible. Comparison with the profiles obtained by chromatography on Cibacron Blue 3-GA and on Green TSK-AF indicates significant differences between the binding properties of the three gels.
Hanahan
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AE
Department of Basic and Clinical Immunology and Microbiology, Medical University of South Carolina, Charleston 29425.
Miribel
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L
Arnaud
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P
D000818:Animals; D001798:Blood Proteins / Q000302:isolation & purification*; D002417:Cattle; D002846:Chromatography, Affinity; D004396:Coloring Agents; D006801:Humans; D007123:Immunoelectrophoresis, Two-Dimensional; D007202:Indicators and Reagents; D011108:Polymers; D013056:Spectrophotometry, Ultraviolet; D014227:Triazines
D001798:Blood Proteins; C053251:Brown Fractogel TSK-AF; D004396:Coloring Agents; D007202:Indicators and Reagents; D011108:Polymers; D014227:Triazines