Through insilico and invitro studies we able to come up with two phyto molecules that are showing considerable DPP4 inhibitory activity. Now we would like to perform DPP8 and DPP9 inhibitory assay to understand the selectivity of our test ligands towards DPP4. we are looking for methodology in this regard.
Following is the paper which explains one methodology, but which we cant execute due to facility concerns.
Establishment of a selective evaluation method for DPP4 inhibitors based on recombinant human DPP8 and DPP9 proteins.
Jinglong Liu, Yi Huan, Caina Li, Minzhi Liu, Zhufang Shenn
Acta Pharmaceutica Sinica B 2014;4(2):135–140
We would like to know any other procedures or if any body working on the same aspect?
Hi
You can try using these commercially available activity assay kits, or just look at these protocols for ideas:
http://bpsbioscience.com/fluorogenic-dpp8-assay-kit-80208
http://bpsbioscience.com/fluorogenic-dpp9-assay-kit-80209
The Fluorogenic Assay Kit is designed to measure activity using purified DPP8 for screening and profiling applications. The key to the Fluorogenic DPP8 Assay Kit is the fluorogenic substrate. Using this kit, only one simple step on a microtiter plate is required for DPP8 reactions. The fluorometric substrate is incubated with a sample containing DPP8 enzyme to produce a fluorophore that can then be measured using a fluorescence reader.
Best of luck
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