By employing a separation-based assay detection platform, researchers can successfully develop challenging assays for lipid-modifying enzymes in a rapid time frame. This approach allows researchers to measure the activity of enzymes with low kinetic activity and enzymes that cannot be analyzed on existing platforms. Compared to conventional plate-based assay technologies, separations allow for quantitation of each component and are less prone to interferences, yielding a greater degree of certainty in the result.
The separation-based approach offers greater flexibility in attaching a tag anywhere on a substrate, which, in turn, speeds development time. Also, separation-based assays provide ratiometric analysis of substrate to product, which allows sensitive detection of low conversion levels, thus ideal for real-time kinetics.
Based on microparallel liquid chromatography (µPLC), Nanostream (www.nanostream.com) developed the Nanostream LD System, a detection platform for assays throughout the screening process—from target characterization through to lead optimization. µPLC miniaturizes liquid chromatography by enabling 24 simultaneous separations and real-time UV and fluorescence detection. By operating in a truly parallel fashion, µPLC allows for the analysis of more samples in less time, which eliminates throughput bottlenecks.
With the Nanostream LD System, researchers can analyze a large number of compounds, increase the number of replicates or conditions used in a study, and reduce solvent consumption and mixed waste generation. Also, the Nanostream LD System integrates with existing workflow and employs commercially available reagents and consumables.
The Nanostream LD System is ideal for developing and screening lipid-modifying enzyme. Method development is simple and iterative with the Nanostream LD System since researchers can optimize conditions for enzyme assays quickly and in parallel, thereby shortening the assay-development time from months to days while achieving higher quality data. By analyzing 24 samples simultaneously, researchers can employ a matrix-based, multiparameter testing approach to define optimal concentrations of buffer constituents, substrates, co-factors, and incubation times.