Dye-affinity purification of transthyretin from an unexploited by-product of human plasma chromatographic fractionation.
Dye-affinity chromatography of human plasma transthyretin on Remazol Yellow GGL-Sepharose from an unexploited by-product of chromatographic fractionation of plasma was optimized for large-scale preparation of a therapeutic product. With this system, transthyretin is only weakly bound to the gel. The residence time on gel and the transthyretin level in the by-product were observed to have no influence on the binding capacity of gel, and the optimum amount of transthyretin to be applied to the gel was found to be 1 g/l of gel. The adsorbent can be used more than ten times. The procedure resulted in the isolation, with a 30% yield with respect to plasma, of an 80% pure protein, which retained its thyroxine-binding capacity. Although the purity is acceptable for substitutive therapy, it can be improved further with a second chromatography on Cibacron Blue-Sepharose.
Regnault
V
V
INSERM U 284, Vandoeuvre-les-Nancy, France.
Rivat
C
C
Vallar
L
L
Stoltz
J F
JF
D000327:Adsorption; D001391:Azo Compounds; D001798:Blood Proteins / Q000032:analysis; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D000431:Ethanol; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D011228:Prealbumin / Q000302:isolation & purification*; D011485:Protein Binding; D012685:Sepharose / Q000031:analogs & derivatives; D012709:Serum Albumin / Q000302:isolation & purification; D013974:Thyroxine / Q000378:metabolism
D001391:Azo Compounds; D001798:Blood Proteins; D007074:Immunoglobulin G; D011228:Prealbumin; D012709:Serum Albumin; C017842:cibacron blue-sepharose; C035187:reactive yellow 13; D000431:Ethanol; D012685:Sepharose; D013974:Thyroxine