[Separation of albumin and IgG by direct chromatographic fractionation of human plasma on DEAE and CM-trisacryl M]. / Préparation de l'albumine et des immunoglobulines G par fractionnement chromatographique direct du plasma humain sur DEAE et 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.