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

Anuradha Subramanian, Peter W. Carr, Clayton V. McNeff

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)15-23
Number of pages9
JournalJournal of Chromatography A
Volume890
Issue number1
DOIs
StatePublished - Aug 18 2000

Fingerprint

Microspheres
Cell culture
Immunoglobulins
Cell Culture Techniques
ethylenediamine
Monoclonal Antibodies
Immunosorbents
Spray drying
zirconium oxide
Electrophoresis
X-Ray Diffraction
Electron Scanning Microscopy
Cations
Assays
Gels
Enzyme-Linked Immunosorbent Assay
X ray diffraction
Scanning electron microscopy
Acids
Enzymes

Keywords

  • Immunoglobulins
  • Monoclonal antibodies
  • Proteins
  • Stationary phases, LC
  • Zirconia

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Use of spray-dried zirconia microspheres in the separation of immunoglobulins from cell culture supernatant. / Subramanian, Anuradha; Carr, Peter W.; McNeff, Clayton V.

In: Journal of Chromatography A, Vol. 890, No. 1, 18.08.2000, p. 15-23.

Research output: Contribution to journalArticle

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