纤维素纤维上CBD-FITC共轭物表面涂层的定量分析。
Quantification of the CBD-FITC conjugates surface coating on cellulose fibres.
作者信息
Pinto Ricardo, Amaral António L, Ferreira Eugénio C, Mota Manuel, Vilanova Manuel, Ruel Katia, Gama Miguel
机构信息
IBB-Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, Universidade do Minho, Campus de Gualtar, 4710-057 Braga, Portugal.
出版信息
BMC Biotechnol. 2008 Jan 9;8:1. doi: 10.1186/1472-6750-8-1.
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).
背景
将纤维素结合结构域（CBD）与异硫氰酸荧光素（FITC）偶联。通过荧光图像分析测定吸附在纤维素纤维上的结合结构域的表面浓度。
结果
对于浓度为60 mg/L的CBD - FITC，Portucel纤维和Whatman纤维的包覆率分别估计为78%和110%。对于饱和CBD浓度，使用Whatman CF11纤维，估计表面浓度为25.2×10⁻¹³ mol/mm²，相当于4个蛋白质单分子层。该结果并不意味着存在多个吸附的蛋白质层。
结论
根据共聚焦显微镜和透射电镜免疫标记分析，证实CBD能够穿透纤维。无定形纤维（磷酸肿胀）上吸附的CBD表面浓度高于结晶度更高的纤维（Whatman CF11和Sigmacell 20）。
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