Encapsulation of Enzymes into PRINT Particles (2008)
Undergraduate: Jesse White
Faculty Advisor: Joseph DeSimone
Department: Chemistry
The imprint lithography method termed PRINT("Particle Replication in Non-wetting Templates") was used to fabricate nanoparticles made of poly(ethylene glycol)400-diacrylate. Given the unique cellular targeting and nanocarrying capabilities of nanoparticles, the ability to infuse enzymes inside the particles was of interest. Enhancement of the current “enzyme replacement therapy” for Gaucher’s Disease was attempted by infusing ?-glucosidase within the nanoparticles and monitoring the enzymatic activity. ?-glucosidase, the enzyme deficient for individuals with Gaucher’s Disease, was encapsulated within the nanoparticles at 0.2 weight percent. The enzyme-substrate activity was measured by an assay that utilized the fluorogenic artificial substrate, 4-methylumbelliferyl-?-D-glucopyranoside. By use of a fluorescent spectrophotometer, comparisons of enzyme-substrate activity of ?-glucosidase free in solution and ?-glucosidase within the nanoparticles yielded an active enzyme concentration estimate of 1.4 nM (0.0003 mg/mL). The activity was also measured in the presence of pronase (a non-specific protease) to determine if enzymes within the nanoparticles were protected from outside digestion mechanisms. When pronase was present in solutions of enzyme-particles, the enzyme-substrate activity decreased by 36.7%. As a result, the enzyme-particles were unable to protect ?-glucosidase from protease digestion. More experiments are needed to determine protection characteristics.