Selection of pH-related parameters in ion-exchange chromatography using pH-gradient operations

T. Ahamed, S. Chilamkurthi, B. Nfor, L. van der Wielen, P. Verhaert, G. Dedem, E. van de Sandt, M.H.M. Eppink, M. Ottens

Research output: Contribution to journalArticleAcademicpeer-review

28 Citations (Scopus)

Abstract

This work demonstrates that the type of ion-exchanger (anion or cation), the mode of operation (bind-and-elute or flow-through), and the operational pH of ion-exchange chromatography (IEX) can be selected in a fast and rational way by analytical pH-gradient IEX operations, thereby eliminating the need for pH scouting or high-throughput screening. The developed approach was applied for the selection of an IEX process for the capture of a monoclonal antibody (MAb) from hybridoma cell culture supernatant (CCS). It was found within a day that MAb can optimally be captured by bind-and-elute mode cation-exchange chromatography (CEX) at pH 4.5 or anion-exchange chromatography (AEX) at pH 7.2 without lowering the salt concentration in the CCS. The performance of both CEX and AEX was predicted to be equal for this particular MAb capture.
Original languageEnglish
Pages (from-to)22-29
JournalJournal of Chromatography. A, Including electrophoresis and other separation methods
Volume1194
Issue number1
DOIs
Publication statusPublished - 2008

Keywords

  • recombinant protein-purification
  • monoclonal-antibody purification
  • liquid-chromatography
  • culture supernatant
  • mass-spectrometry
  • optimization
  • separation
  • retention
  • capture
  • design

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