OBJECTIVE: Enzymatic fingerprinting of key enzymes of glucose metabolism is a valuable analysis tool in cell physiological phenotyping of plant samples. Yet, a similar approach for mammalian cell line samples is missing. In this study, we applied semi-high throughput enzyme activity assays that were originally designed for plant samples and tested their feasibility in extracts of six frequently used mammalian cell lines (Caco2, HaCaT, C2C12, HEK293, HepG2 and INS-1E). RESULTS: Enzyme activities for aldolase, hexokinase, glucose-6-phosphate dehydrogenase, phosphoglucoisomerase, phosphoglucomutase, phosphofructokinase could be detected in samples of one or more mammalian cell lines. We characterized effects of sample dilution, assay temperature and repeated freeze-thaw cycles causing potential biases. After careful selection of experimental parameters, the presented semi-high throughput methods could be established as useful tool for physiological phenotyping of cultured mammalian cells.

Department of Science and Environment Roskilde University Universitetsvej 1 4000 Roskilde Denmark

Global Change Research Institute CAS Drásov Czech Republic

Section for Crop Sciences Department of Plant and Environmental Sciences Copenhagen Plant Science Centre University of Copenhagen Højbakkegårds Allé 13 2630 Taastrup Denmark

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Simple semi-high throughput determination of activity signatures of key antioxidant enzymes for physiological phenotyping