I'm currently working on my graduation project on the anticancer potential of E. coli L-asparaginase enzyme on mcf-7 along with other chemotherapy drugs. Previous references did prove cytotoxic effect with low concentrations but it hardly gave me a suitable IC50 when I used around 500IU/ml. Even in combination with other drugs, it didn't make much difference which is not what the references have stated. Could there be something wrong with the enzyme I was provided with? Or could it be the media I used? I used RPMI as cell culture media but I found some papers saying it would be better to use an L-glutamine and asparagine free media for the experiment. Thank you in advance.
Was the E.coli enzyme well purified? How much endotoxin comes with the enzyme? Check the enzyme preparation for lipopolysaccharides (LPS). You can buy commercial LPS tests.
Obviously you need to see that the medium you have chosen shall be devoid of asparagine and glutamine. Most readymade commercial media contain both of these. Thus there shall be greater need to add more of asparaginase enzyme.
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