Diffusion and transfer of antibody proteins from a sugar-based hydrogel |
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Authors: | Michael A. Markowitz David C. Turner Brett D. Martin Bruce P. Gaber |
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Affiliation: | (1) Laboratory for Molecular Interfacial Interactions, Code 6930, Naval Research Laboratory, 20375 Washington, D. C. |
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Abstract: | Diffusion of antibody protein from hydrogel films and hydrogel encapsulated in a microcapillary was studied. Thin hydrogel films were formed by crosslinking 6-acryloyl-B-O-methylgalactoside withN,N’-methylene-bis-acrylamide and the diffusive transport of monoclonal antimouse IgG-FITC into and out of the hydrated films was measured. Diffusion coefficients in 2 and 4% crosslinked hydrogel films were measured. The measured diffusion constants determined for IgG in both the 2 and 4% hydrogel films were comparable to the free diffusion of IgG in bulk water (D mean ∼ 10-7cm2/s). In addition, 2% crosslinked hydrogels were prepared in a capillary tube and the transport of antimouse IgG-FITC into and out of the hydrated hydrogel was measured. Kinetic analysis indicated that the protein transport through the capillary hydrogel was faster than would be expected for a simple diffusion process. Finally, by utilizing the diffusion of antibody from the capillary hydrogel, transfer of antibody to a silica surface was demonstrated. A capillary hydrogel loaded with antimouse IgG-FITC was used to transfer the protein to a silica surface forming a 30-μm spot of antibody, which was imaged using fluorescence microscopy. These results may lead to the development of a nonlithographic method of patterning antibodies on surfaces for use in integrated microimmunosensors. |
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Keywords: | Hydrogel multianalyte sensor immunosensor device miniaturization diffusion transfer capillary antibody |
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