The gel forming characteristics of alginate in the presence of calcium ions and further crosslinking with poly-L-lysine led to the formation of sponge-like nano- and microparticles. The particle size was varied by adjusting the final concentrations of and proportions between the components. The region for particle formation was from 0.04 to 0.08% (w/v) of alginate in the final formulation, the change from the nm to microm size range occurred at a concentration of approx. 0.055% (w/v). Oligonucleotide-loaded microparticles were prepared by two different methods, either by absorption of the drug into the crosslinked polymeric matrix or by incorporation of an oligonucleotide/poly-L-lysine complex into a calcium alginate pre-gel. The release of oligonucleotide from microparticles prepared by the first method was higher. The addition of increasing amounts of poly-L-lysine resulted in larger particles, higher oligonucleotide loading and slower drug release. An increase in the final solid content of the formulation led to larger particles, especially with high concentrated calcium alginate pre-gels. Microparticles based on alginate and poly-L-lysine are potential carriers for antisense oligonucleotides.