Diffusion and transfer of antibody proteins from a sugar-based hydrogel.
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 with N,N'-methylene-bis-acrylamide and the diffusive transport of monoclonal antimouse IgG-FITC into and out of the hydrate 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 (Dmean approximately 10(-7) cm2/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-micron 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.
Markowitz
M A
MA
Laboratory for Molecular Interfacial Interactions, Naval Research Laboratory, Washington, D.C. 20375, USA.
Turner
D C
DC
Martin
B D
BD
Gaber
B P
BP
D000178:Acrylamides; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D002197:Capillary Action; D003432:Cross-Linking Reagents; D004058:Diffusion; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000378:metabolism; D008756:Methylgalactosides; D051379:Mice; D008856:Microscopy, Fluorescence; D011108:Polymers
D000178:Acrylamides; D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D008756:Methylgalactosides; D011108:Polymers; C109801:hydrogel film; C021221:N,N'-methylenebisacrylamide; D016650:Fluorescein-5-isothiocyanate