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Biological activity of lysozyme after entrapment in poly(d,l-lactide-co-glycolide)-microspheres
1Uppsala University, Department of Pharmaceutics, Sweden. raouf.ghaderi@galenik.uu.se
Purpose:
The purpose of this study was to investigate the process of preparing microspheres for maximising entrapment efficiently (EE) and retained biological activity (RBA) of peptides and proteins.
Methods:
A controlled-release formulation based on poly(d,l-lactide-co-glycolide) was designed and produced using a small-scale double emulsion method. These PLG microspheres contained a model peptide, lysozyme. The retained bioactivity of the incorporated lysozyme was determined by bacterial assay. The size distributions and the morphology of the microspheres were characterized.
Results:
The RBA and EE improved when the PLG concentration in the organic phase of the emulsion was increased. A high lysozyme concentration in the inner water phase of the emulsion resulted in decreased EE and an increase in the proportion of fragmented particles. The RBA of lysozyme in the microspheres varied between 30 and 80% with changes to the process.
Conclusions:
The study shows that the RBA of lysozyme in PLG microspheres is strongly dependent on the experimental conditions for preparing the microspheres. Measurement of the EE alone, without the RBA is insufficient to evaluate the efficacy of the designed delivery system.