This document introduces the characteristics of Agilent RapidFire/MS system for high-throughput screening intractable targets, the system uses label-free native analyte detection, eliminates the need for cumbersome labeling methods and surrogate substrates that may not accurately represent the native reaction, enables traditionally low-throughput, intractable assays to be converted into high-throughput assays processed at speeds approaching plate-based optical methods.
This document describes an application note detailing the analysis of UDP-3-0-(R-3-hydroxymyristoyl)-N-Acetylglucosamine Deacetylase (LpxC) using Agilent RapidFire/MS Systems. The text explains that traditional assays for this essential enzyme often require cumbersome labeling methods or imperfect surrogate substrates, which can lead to false positives or inaccurate kinetics.
The RapidFire/MS method offers a fast, sensitive, and reliable alternative by employing mass spectrometry to directly detect the native product without the need for surrogate substrates. The document presents data illustrating enzyme concentration linearity, standard enzyme kinetics, and results from a screening campaign against a random compound library, highlighting the system's ability to uncover novel inhibitors.
| Sample processing speed | 6-10 seconds |
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It eliminates the need for cumbersome labeling methods and surrogate substrates by directly detecting the native product.
It employs a solid phase extraction (SPE) sample cleanup step directly coupled to MS detection.