Quantification of the CBD-FITC conjugates surface coating on cellulose fibres

BMC Biotechnol. 2008 Jan 9:8:1. doi: 10.1186/1472-6750-8-1.

Abstract

Background: Cellulose Binding Domains (CBD) were conjugated with fluorescein isothiocyanate (FITC). The surface concentration of the Binding Domains adsorbed on cellulose fibres was determined by fluorescence image analysis.

Results: For a CBD-FITC concentration of 60 mg/L, a coating fraction of 78% and 110% was estimated for Portucel and Whatman fibres, respectively. For a saturating CBD concentration, using Whatman CF11 fibres, a surface concentration of 25.2 x 10-13 mol/mm2 was estimated, the equivalent to 4 protein monolayers. This result does not imply the existence of several adsorbed protein layers.

Conclusion: It was verified that CBDs were able to penetrate the fibres, according to confocal microscopy and TEM-immunolabelling analysis. The surface concentration of adsorbed CBDs was greater on amorphous fibres (phosphoric acid swollen) than on more crystalline ones (Whatman CF11 and Sigmacell 20).

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cellulases / genetics
  • Cellulases / metabolism*
  • Cellulose / metabolism*
  • Cellulose / ultrastructure
  • Fluorescein-5-isothiocyanate / metabolism*
  • Fluorescence
  • Microscopy, Confocal
  • Microscopy, Electron, Transmission
  • Protein Binding
  • Protein Structure, Tertiary

Substances

  • Cellulose
  • Cellulases
  • Fluorescein-5-isothiocyanate