EFFECT OF POLYSORBATE-80 CONCENTRATION ON G-CSF FORMULATION USING LIQUID CHROMATOGRAPHY
Objective: The aim of the study was to find the dual effects of polysorbate-80 concentration on oxidation and aggregation in the formulation of granulocyte colony stimulating factor.
Methods: The effect of polysorbate-80 at different concentration was monitored and analyzed using high performance liquid chromatography. Oxidative degradation and aggregate generation was studied using reverse phase and size exclusion chromatography method respectively.
Results: With increase in concentration of polysorbate-80 the amount of oxidation as well as aggregate formation increases in a concentration dependent manner. The aggregates present at higher concentration of polysorbate-80 formulation are not found with low concentration or without polysorbate-80. This result shows that higher concentration of Tween-80 force the formation of oligomers and leads to increased level of oxidation of granulocyte colony stimulating factor.
Conclusion: The dual effect of aggregation and oxidation of polysorbate-80 on the recombinant granulocyte colony stimulating factor indicate that during formulation development studies it is crucial to evaluate the amount of polysorbate-80 to be used in the formulation.
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