Advertisement

Chromatographia

, Volume 58, Issue 5–6, pp 365–373 | Cite as

Ion-Exchange Chromatographic Supports Obtained by Formation of Polyelectrolyte Multi-Layers for the Separation of Proteins

  • M.-C. Millot
  • T. Debranche
  • A. Pantazaki
  • I. Gherghi
  • B. Sébille
  • C. Vidal-Madjar
Article

Summary

High performance liquid chromatography (HPLC) was used to study the mechanism of formation of polyelectrolyte multilayers on porous silicas. The coatings were produced by alternating the adsorption of positively and negatively charged polymers. The stationary phases formed by adsorbing a single layer, double layers and triple layers were tested by studying the elution behavior of model proteins. The double polymer coating was achieved by adsorbing first a polycation such as hexadimethrine bromide (HB) on the HPLC silica support and then a polyanion such as dextran sulfate (DS) on the cationic layer formed. The retention properties of this support are mainly those of a cation exchanger as the negatively charged proteins were strongly retained while positively charged ones were weakly adsorbed. This work demonstrated the importance of the first underlying layer as the retention behavior of proteins was greatly affected by the properties of this coating. The triple polymer coating was achieved by adsorbing the polycation (HB) on the double layer coating (HB-DS). Its retention behavior was that of an anion exchange support. The HB-DS stationary phase displayed good chromatographic performances, with an adsorbed layer relatively stable. The polyelectrolyte multilayer coating procedure was useful to easily synthesize cation-exchange supports for the separation of basic proteins.

Key Words

Ion-exchange chromatography Polyelectrolyte layers Silica coatings Proteins 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Copyright information

© Friedr. Vieweg&Sohn Verlagsgesellschaft mbH 2003

Authors and Affiliations

  • M.-C. Millot
    • 1
  • T. Debranche
    • 1
  • A. Pantazaki
    • 2
  • I. Gherghi
    • 3
  • B. Sébille
    • 1
  • C. Vidal-Madjar
    • 1
  1. 1.Laboratoire de Recherche sur les PolyméresCNRS-Université Paris Val de MarneThiaisFrance
  2. 2.Laboratory of BiochemistryAristotle University of ThessalonikiGreece
  3. 3.Laboratory of Analytical ChemistryAristotle University of ThessalonikiThessalonikiGreece

Personalised recommendations