5 years ago

Visualization and Quantification of IgG Antibody Adsorbed at the Cellulose–Liquid Interface

Visualization and Quantification of IgG Antibody Adsorbed at the Cellulose–Liquid Interface
Stephen A. Holt, Christopher J. Garvey, Gil Garnier, Vikram Singh Raghuwanshi, Warren J. Batchelor, Peter J. Holden, Jielong Su
Quantification of adsorbed biomolecules (enzymes, proteins) at the cellulose interface is a major challenge in developing eco-friendly biodiagnostics. Here, a novel methodology is developed to visualize and quantify the adsorption of antibody from solution to the cellulose–liquid interface. The concept is to deuterate cellulose by replacing all nonexchangeable hydrogens from the glucose rings with deuterium in order to enhance the scattering contrast between the cellulose film surface and adsorbed antibody molecules. Deuterated cellulose (DC) was obtained from bacterial (Gluconacetobacter xylinus strain) cellulose, which was grown in heavy water (D2O) media with a deuterated glycerol as a carbon source. For comparison, hydrogenated cellulose (HC) was obtained from cellulose acetate. Both HC and DC thin films were prepared on silicon substrate by spin coating. X-ray reflectivity (XR) shows the formation of homogeneous and smooth film. Neutron reflectivity (NR) at the liquid/film interface reveals swelling of the cellulose film by a factor of 2–3× its initial thickness. An Immunoglobulin G (IgG), used as a model antibody, was adsorbed at the liquid–solid interface of cellulose (HC) and deuterated cellulose (DC) films under equilibrium and surface saturation conditions. NR measurements of the IgG antibody layer adsorbed onto the DC film can clearly be visualized, in sharp contrast in comparison to the HC film. The average thickness of the IgG adsorbed layer onto cellulose films is 127 ± 5 Å and a partial monolayer is formed. Visualization and quantification of adsorbed IgG is shown by large difference in scattering length density (SLD) between DC (7.1 × 10–6 Å–2) and IgG (4.1 × 10–6 Å–2) in D2O, which enhanced the scattering contrast in NR. Quartz crystal measurements (QCM-D) were used as a complementary method to NR to quantify the adsorbed IgG over the cellulose interface.

Publisher URL: http://dx.doi.org/10.1021/acs.biomac.7b00593

DOI: 10.1021/acs.biomac.7b00593

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