Chitosan/tripolyphosphate (CHIT/TPP) and chitosan/tripolyphosphate/chondroitin sulfate (CHIT/TPP/CHS) core-shell type microspheres were prepared by polyelectrolyte complexation in order to develop a biocompatible matrix for drug delivery. The continual method using a multi-loop reactor under sterile conditions was applied for microsphere preparation. All the types of microspheres produced were spherical in shape and had a porous structure. The mechanical resistance of the microspheres increased in the presence of CHS as the second polyanion, which toughened the microsphere shell structure. For a drug release application, the process of microsphere preparation was modified by dissolving ofloxacin (OFL), the fluoroquinolone antibiotic, in CHIT solution before complex formation. This study shows the difference in OFL release comparing the microspheres CHIT/TPP and CHIT/TPP/CHS and implies the potential to control this process.