Identification of the mass transfer mechanisms involved in the transport of human immunoglobulin-G in N,N,N′,N′- ethylenediaminetetramethylenephosphonic acid-modified zirconia

Sabyasachi Sarkar, Peter W. Carr, Anu Subramanian

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Zirconia particles modified with N,N,N′,N′- ethylenediaminetetramethylenephosphonic acid (EDTPA), further referred to as r_PEZ, were studied as a support material for use in chromatography. Our previous studies have demonstrated the utility of r_PEZ in the separation of immunoglobulins from biological fluids. In the present study we sought to understand the underlying factors and identify the rate-limiting mechanisms that govern the transport of biomolecules in r_PEZ. Pulse injection techniques were used to elucidate the individual mass transfer parameters. Elution profiles obtained under retained and unretained conditions were approximated by the Gaussian equation and the corresponding HETP contributions were estimated. The dependence of the HETP values on incremental salt concentration in the mobile phase was determined. Resulting data in conjunction with the equations outlined in literature were used to estimate the theoretical number of transfer units for the chromatographic separation process. Our results indicate that surface diffusion probably plays a minor role; however pore diffusion was established to be the rate limiting mechanism for immunoglobulin G adsorption to r_PEZ. The HETP based methodology may be used to estimate the rate limiting mechanisms of mass transfer for any given chromatographic system under appropriate conditions.

Original languageEnglish (US)
Pages (from-to)124-131
Number of pages8
JournalJournal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
Volume821
Issue number2
DOIs
StatePublished - Jul 25 2005

Keywords

  • HETP
  • Immunoglobulin
  • Pseudo-affinity matrix
  • Pulse-injection
  • Zirconia

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Cell Biology

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