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A new miniaturized micro sequential injection coupled with the lab-on-valve (mu SIA-LOV) technique was full-blown to perform inhibitory studies on dihydrofolate reductase (DHFR). The system was used to evaluate the DHFR inhibition activity of metal-based anticancer compounds. The metal complexes exhibited IC50 values in the range 1.3 +/- 0.3-108 +/- 7 mu M, with half of the complexes lying in the low mu M range, i.e., 1.3 +/- 0.3-4.4 +/- 0.2 mu M. For comparison, methotrexate (MTX), a known inhibitor of DHFR, has an IC50 value of 0.38 +/- 0.06 mu M. The mu SIA-LOV is a versatile, robust, rapid, and easy to operate, allowing automated determination of DH F R inhi-ition. Moreover, the automated system requires very little sample (approximately 40 mu L per analysis), uses minimal reagents (5 times less than the batch procedure used), and generates very little waste (around 1.2 mL per analysis) compared with batch methods, considerably reducing costs.
Henning Paul-Julius Stahlberg, Dongchun Ni