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 microm) 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.
Subramanian
A
A
University of Minnesota, Department of Biosystems and Agricultural Engineering, St. Paul 55108, USA.
Carr
P W
PW
McNeff
C V
CV
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000302:isolation & purification; D018929:Cell Culture Techniques; D002853:Chromatography, Liquid / Q000379:methods*; D003470:Culture Media / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007136:Immunoglobulins / Q000302:isolation & purification*; D051379:Mice; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D016062:Porosity; D014961:X-Ray Diffraction; D015040:Zirconium / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D003470:Culture Media; D007136:Immunoglobulins; D015040:Zirconium; C028541:zirconium oxide