High-throughput assay platform development

RNA modification often involves chemical changes such as the addition of a methyl group or removal of a phosphate group that were traditionally studied using radiometric methods. These previous methods are tedious and low in resolution power. LC-MS/MS techniques, on the other hand, are a powerful tool for direct detection of these subtle chemical manipulations performed by RNA modifying enzymes. We have established methods to interface complex LC-MS/MS methods with enzymology experiments to probe the reactant composition of enzymatic reactions at high-throughput.

In this example, 96 gene products are tested for their activity in converting a substrate into three different products. Our high-throughput (HT) assay platform allows LC-MS/MS analysis of these reactions in a 96-well scale.

In another example, 96 gene products are tested for their activity to convert four different substrates into their respective products. Our HT LC-MS/MS assay platform enables relative quantification of each of the substrates and products in a multiplex format.