A method suitable for the isolation of monoclonal antibodies (MAbs) on novel zirconia microspheres (20-30 μm) 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(methylenephosphonic) 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.
Bibliographical noteFunding Information:
This work was performed through a grant from the National Institutes of Health (NIH PR 1R43-GM58354-01). The technical assistance of Lorinda Zigan is truly appreciated.
- Monoclonal antibodies
- Stationary phases, LC