Interaction of immunoglobulin G with modified chitosan

Anuradha Subramanian, Jennifer Hommerding

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

Chitosan beads were modified to include a spacer arm and end-capped with a caboxyethyl-group containing anionic ligand to generate a pseudo-bioaffinity support, further referred to as Ligosep Alpha® or LMCB. To better understand the force of interaction between the immunoglobulin G (IgG) and the chitosan-based pseudo-affinity support, the equilibrium dissociation constant (Kd) was determined by static binding isotherms, as a function of temperature, and by frontal analysis at different linear velocities. The maximum static binding capacity (Qmax) was found to be in the range of 60-70 mg IgG per mL of gel, and unaffected by temperature. The adsorption rate constant (ka) was determined by a split-peak approach to be between 46 and 404 L mol-1 sec-1 depending on the linear velocity. A governing empirical equation relating ka to the linear velocity (u) was deduced to be ka = 44.4(u) + 9.3. The kinetic parameters of the chromatographic system indicated a good capacity but a low adsorption rate constant. LMCB was found to preferentially interact with the Fab and the F(ab)2 region of the HIgG molecule. This gives the further potential of adjusting the process for purifying specific paratopes under gentle chromatographic conditions.

Original languageEnglish (US)
Pages (from-to)2671-2688
Number of pages18
JournalJournal of Liquid Chromatography and Related Technologies
Volume27
Issue number17
DOIs
StatePublished - Oct 21 2004

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Keywords

  • Chitosan
  • Immunoglobulin G

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Pharmaceutical Science
  • Clinical Biochemistry

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