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Quantification of the CBD-FITC conjugates surface coating on cellulose fibres
Publication . Pinto, Ricardo; Amaral, António L.; Ferreira, Eugénio C.; Mota, Manuel; Vilanova, Manuel; Ruel, Katia; Gama, Miguel
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/mm(2) 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).
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Fundação para a Ciência e a Tecnologia
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SFRH
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SFRH/BD/6934/2001
