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Background . L-asparaginase is effective in treating canine and feline lymphoma, however chemotherapy poses a significant financial cost to veterinary clients, limiting therapy for many pets. Single dose vials result in significant drug wastage, and drug shortages limit consistent availability for pets.  Hypothesis .  E. coli -derived asparaginase retains enzymatic and antineoplastic activity in canine and feline lymphoma cells after cold storage.  Methods .  E. coli -derived asparaginase was cold-stored: refrigeration (7–14 days) and freezing (14 days–six months, one to three freeze/thaw cycles). Enzymatic activity of asparaginase was measured via a modified asparagine assay. Effects of cold-stored asparaginase on cell proliferation and cytotoxicity were measured in feline (MYA-1, F1B) and canine (17–71, OSW) lymphoma cells.  Results . Cold-stored  E. coli -derived asparaginase retains antineoplastic activity in all four cell lines tested. Cold-stored  E. coli -derived L-asparaginase depletes asparagine and retains enzymatic activity. Duration of refrigeration, duration of freezing, and number of freeze-thaw cycles have minimal effect on asparaginase enzyme activity.  Conclusions and Clinical Importance . This study establishes a scientific basis for long-term cold storage of reconstituted  E. coli -derived asparaginase that may result in better utilization of limited drug resources and improve financial feasibility of  E. coli -derived asparaginase as a therapeutic option for pets with lymphoma.

E. coli-Derived L-Asparaginase Retains Enzymatic and Cytotoxic ActivityIn Vitrofor Canine and Feline Lymphoma after Cold Storage