Electrophoretic extraction and analysis of DNA from chitosan or poly-L-lysine-coated alginate beads.
Alginate beads containing entrapped DNA were produced using both external and internal calcium sources, and coated with chitosan or poly-L-lysine membranes. The beads were assayed with DNase nuclease to determine formulation conditions offering the highest level of DNA protection from nucleic acid hydrolysis, simulating gastrointestinal exposure. A method was developed to extract and assay intracapsular DNA through a modified agarose electrophoresis system. Both external and internally gelled beads were permeable to DNase (Mw = 31 kDa), indicated by the absence of DNA after nuclease exposure. At low levels of DNase exposure, coated high guluronic content alginate beads offered a higher level of DNA protection compared with coated beads with low guluronic alginate. No apparent correlation was found with chitosan membrane molecular weight and degree of deacetylation; however, increasing poly-L-lysine molecular weight appeared to increase DNase exclusion from beads. At elevated levels of DNase exposure, DNA hydrolysis was evident within all coated beads with the exception of those coated with the highest molecular weight poly-L-lysine (Mw = 197.1 kDa), which provided almost total nuclease protection. Optimal combination then for DNA protection from nucleases is a high guluronic alginate core, coated with high molecular weight poly-L-lysine.
Quong
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Department of Chemical Engineering, McGill University, 3480 University Street, Montréal, Québec, H3A 2A7, Canada.
Neufeld
R J
RJ