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P9005
Real-time Reaction Monitoring & Enzyme Kinetics Measurements on the ExpressLC platform
Presenter Don Arnold, Eksigent Technologies, USA
Additional Authors: Maxim Shusteff, H.-Y. Cheng, Christopher Hoyle, David Wyrick, Phillip H. Paul and Don W. Arnold
Microscale liquid chromatography (uLC) systems, when fabricated using high-precision flow control components offer the ability to carry out fast, repeatable separations on very small amounts of sample using with standard LC materials and methods. As such, we find that the Eksigent ExpressLC is particularly useful for carrying out LC-based biological assays where reagents are often precious. When combined with system’s novel sample introduction methods, the ExpressLC’s utility is extended further. We demonstrate a novel approach for determining enzyme kinetics and real-time reaction rate monitoring using this system. Using only a very small sample quantity (~10uL or less; or attomole-range reagent consumption), the total assay can be carried out.  It offers the flexibility to carry out label-free assays using absorbance detection or more specific assays with sensitive fluorescence detection. The system’s simple configuration facilitates the collection of real-time quantitative information as the assay progresses, at turnover rates as low as 1%.