Application of dye-ligant chromatography to the isolation of alpha-1-proteinase inhibitor and alpha-1-acid glycoprotein.
Various Cibacron blue F3G-A substituted Sephadex G-100 gels which differ in the density of the bound dye were investigated for their applicability in the affinity chromatography of human serum proteins. Protein adsorption was found to depend strongly on the density of the covalently attached dye and on the pH of the applied buffer. A high degree of dye substitution of the gel caused binding of most of the serum proteins. Only a small number of proteins were found to appear in the breakthrough fraction. On this basis a simple and relatively mild procedure for the isolation of homogeneous alpha-1-proteinase inhibitor and alpha-1-acid glycoprotein from human serum was developed. Isolation of alpha-1-acid glycoprotein succeeded in only a single step, whereas that of alpha-1-proteinase inhibitor required two additional steps: a prior ammonium sulphate precipitation and a chromatographic step on DEAE-cellulose at pH 6.5.
D001798:Blood Proteins / Q000032:analysis* / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods; D006801:Humans; D009961:Orosomucoid / Q000302:isolation & purification*; D000515:alpha 1-Antitrypsin
D001798:Blood Proteins; D009961:Orosomucoid; D000515:alpha 1-Antitrypsin