Use of spray-dried zirconia microspheres in the separation of immunoglobulins from cell culture supernatant

A method suitable for the isolation of monoclonal antibodies (MAbs) on novel zirconia microspheres (20-30 mu m) is described. Zirconia microspheres were generated by spray drying colloidal zirconia. Spray-dried zirconia microspheres were further classified and characterized by X-ray diffraction, BET porosimetry and scanning electron microscopy. Spray-dried zirconia microspheres were modified with ethylenediamine-N,N'-tetra(methylenephosphinic) acid (EDTPA) to create a cation-exchange chromatographic support. The chromatographic behavior of a semi-preparative column packed with EDTPA-modified zirconia microspheres was evaluated and implications for scale-up are provided. EDTPA-modified zirconia microspheres were further used to purify MAbs from cell culture supernatant. Analysis by enzyme linked immunosorbent assay and gel electrophoresis demonstrate that MAbs can be: recovered from a cell culture supernatant at high yield (92-98%) and high purity (>95%) in a single chromatographic step. (C) 2000 Elsevier Science B.V. All rights reserved.