Preparation, characterization, and process performance of composite fibrous adsorbents as cation exchangers for high throughput and high capacity bioseparations

J Chromatogr B Analyt Technol Biomed Life Sci. 2012 Aug 15:903:14-22. doi: 10.1016/j.jchromb.2012.06.027. Epub 2012 Jun 29.

Abstract

Fibrous materials are proposed as novel chromatographic supports depicting high throughput and high product capacity. In this work, a composite fiber harboring strong cation-exchange moieties has been investigated. Such materials were characterized by a plethora of physical methods including degree of swelling (DS), scanning electron microscope (SEM), confocal laser scanning microscopy (CLSM), and Fourier transform infrared spectroscopy-attenuated total reflection (FTIR-ATR). The composite showed a high degree of grafting (∼30%) and exhibited a high swelling ratio (∼300%). Moreover, homogenous grafting and the development of an internal (functional) hydrogel were observed. The fibrous adsorbent was packed utilizing a designed "double roll" supported-structure and subsequently tested for packing efficiency and chromatography performance. The mentioned system showed similar packing efficiency of height equivalent to a theoretical plate (HETP) value and higher permeability coefficient (0.92 × 10⁻⁷ cm²) than commercial resins. Experimentally determined Peclet number (Pe) values were within the range 60-90, suggesting a close-to-plug-flow condition. Total ionic capacity of the fibrous adsorbent was determined by the transition pH method. A capacity of 6.5 mequiv./g was obtained. Moreover, a high dynamic binding capacity for lysozyme was found to be 283 mg/g. On the other hand, a bed of randomly packed fiber also demonstrated high-resolution ability when a mixture of model protein was utilized to that end. Resolution was maintained at high flow rates (up to 900 cm/h) and utilizing shorter gradient development routines. Direct sequestration of a model protein (lysozyme) was also possible from an artificial mixture containing 1.5% yeast homogenate. Summarizing, the composite fibrous adsorbents exhibited superior performance during early protein capture and intermediate-resolution applications.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adsorption
  • Cation Exchange Resins / chemistry*
  • Cations
  • Chromatography, Ion Exchange / instrumentation*
  • High-Throughput Screening Assays / instrumentation*
  • Hydrogel, Polyethylene Glycol Dimethacrylate / chemistry
  • Hydrogen-Ion Concentration
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / metabolism
  • Microscopy, Confocal
  • Muramidase / chemistry
  • Muramidase / metabolism
  • Permeability
  • Pressure
  • Protein Binding
  • Spectroscopy, Fourier Transform Infrared

Substances

  • Cation Exchange Resins
  • Cations
  • Immunoglobulin G
  • Hydrogel, Polyethylene Glycol Dimethacrylate
  • Muramidase