[Separation of albumin and IgG by direct chromatographic fractionation of human plasma on DEAE and CM-trisacryl M].
We described a continuous chromatographic process using gel-filtration on Ultrogel AcA 202 and ion exchange chromatography on DEAE and CM-Trisacryl M. The human plasma cryosupernatant is firstly desalted on Ultrogel AcA 202 equilibrated in 0.025 M Tris-HCl buffer pH 8.8. containing 0.035 M NaCl and, then, applied to a column of DEAE-Trisacryl M. By these steps, we have obtained following results: - the IgG are not adsorbed and obtained directly with a minimal purity of 99.5%; - a fraction containing alpha2 and beta2 globulin (such as transferrin is removed by elution with 0.025 M Tris-HCl, buffer pH 8.5 containing 0.075 M Na Cl; - crude albumin (purity of 70%) is eluted with 0.025 M Tris-HCl, buffer pH 7.5 containing 0.15 M NaCl. The prepurified albumin is equilibrated on Ultrogel AcA 202 (0.04 M acetate buffer, pH 4.2) and, then, applied to a CM-Trisacryl M column: - the non adsorbed fraction contains alpha and beta-globulins; - a 98% pure albumin is removed by elution with 0.1 M acetate buffer pH 5; - the more adsorbed protein fraction are removed by elution with 0.1 M acetate buffer, pH 4.2. All the chromatographic steps can be completed within 7 h. The purity of the final products fulfills the Pharmacopeia requirements. The characteristics of this plasma protein fractionation process are discussed related to the properties of the new ion exchangers Trisacryl.
D000180:Acrylic Resins; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange / Q000379:methods; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification*; D007475:Ion Exchange Resins; D012709:Serum Albumin / Q000302:isolation & purification*
D000180:Acrylic Resins; D007074:Immunoglobulin G; D007475:Ion Exchange Resins; D012709:Serum Albumin; C034203:CM-trisacryl M
Préparation de l'albumine et des immunoglobulines G par fractionnement chromatographique direct du plasma humain sur DEAE et CM - Trisacryl M.