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Fabrication and Characterization of Polyelectrolyte Microparticles with Protein

N. G. Balabushevich and N. I. Larionova*

Faculty of Chemistry, Lomonosov Moscow State University, Moscow 119992, Russia; fax: (7-095) 939-5417; E-mail: nilar@enzyme.chem.msu.ru

* To whom correspondence should be addressed.

Received December 8, 2003; Revision received December 29, 2003
The incorporation of proteins into microparticles fabricated by layer-by-layer adsorption of oppositely charged polyelectrolytes (dextran sulfate and protamine) on protein microaggregates was studied. Microaggregates with insulin were prepared by two different techniques: 1) formation of insoluble polyelectrolyte complex consisting of insulin and dextran sulfate (aggregate size of 7-20 µm), or 2) salting out of insulin from solution by sodium chloride (aggregate size of 5-13 µm). Microparticles varying in the number of cycles (from 1 to 8) of polyelectrolyte adsorption on protein aggregates were examined and compared. Morphology of the microparticles was studied by scanning electron and optical microscopy. It was shown that polyelectrolyte microparticles retained the shape and dimensions of the initial protein aggregates used as a template. Ultrasonication of microparticles obtained using salted out protein aggregates resulted in the formation of stable nanoparticles (100-200 nm). Regulation of protein release from the microparticles of both types by varying the number of polyelectrolyte adsorption cycles and pH of the medium was demonstrated. Insulin not bound to polyelectrolytes was released from the microparticles at pH values between 6 and 8, which corresponds to the pH of the human small intestine and ileum.
KEY WORDS: proteins, polyelectrolytes, polyelectrolyte microparticles, dextran sulfate, protamine, insulin