Optimization of pressure-flow limits, strength, intraparticle transport and dynamic capacity by hydrogel solids content and bead size in cellulose immunosorbents

Jeffrey A. Kaster, Willer de Oliveira, Wolfgang G. Glasser, William H. Velander

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

The design of existing beaded adsorbent materials for column-mode protein purification has emphasized the impact of diffusional transport phenomena upon adsorbent capacity. A design model is presented here that optimizes molecular accessibility of proteins relative to the mechanical stability at low operating pressures by manipulation of size and solids content for uncross-linked cellulose beads. Cellulose beads of several different sizes ranging from about 250 to 1000 μm diameter and having different solids contents were evaluated. Solids content of greater than about 9% cellulose greatly reduced the permeability of large proteins such as thyroglobulin and β-amylase into the beaded matrix at bead contacting times of about 5 and 50 s. Furthermore, the amount of permeation at 3% solids content by thyroglobulin at bead contacting times of about 5 s was about tenfold larger than predicted by diffusion models using the binary diffusivity in a purely aqueous continuum. The utility of a low solids content, large bead cellulose support was shown with immobilized IgG (Mr 155 kDa) capturing recombinant human Protein C (Mr 62 kDa). A 1000 μm diameter beaded cellulose immunosorbent having 3% solids content gave equivalent capacity to a 140 μm diameter beaded, cross-linked agarose support containing 4% solids. In contrast to the smaller diameter, cross-linked beaded agarose, the low solids content beaded cellulose benefitted from greater physical stability due to more optimal pressure-flow characteristics imparted by large bead size.

Original languageEnglish (US)
Pages (from-to)79-90
Number of pages12
JournalJournal of Chromatography A
Volume648
Issue number1
DOIs
StatePublished - Oct 1 1993

Fingerprint

Immunosorbents
Hydrogel
Cellulose
Pressure
Thyroglobulin
Sepharose
Adsorbents
Proteins
Amylases
Protein C
Recombinant Proteins
Mechanical stability
Permeability
Distillation columns
Immunoglobulin G
Permeation
Purification

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

Cite this

Optimization of pressure-flow limits, strength, intraparticle transport and dynamic capacity by hydrogel solids content and bead size in cellulose immunosorbents. / Kaster, Jeffrey A.; de Oliveira, Willer; Glasser, Wolfgang G.; Velander, William H.

In: Journal of Chromatography A, Vol. 648, No. 1, 01.10.1993, p. 79-90.

Research output: Contribution to journalArticle

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