Final ProgramCPSA 2011
Science and Technology Coming Together to Make a Difference
October 3 - 6, 2011
Bucks County Sheraton Hotel
Langhorne, PA
Poster Abstract #31
High Throughput Screening for Lead Discovery using Multiplexed LC with High Resolution Accurate Mass LC/MS
Thermo Fisher Scientific, 355 River Oaks Parkway, San Jose, CA, 95134, USA
High Throughput Screening is a valuable part of identifying new leads and directing drug design in pharmaceutical research. The ability to quickly perform large numbers of analyses is critical to the success of any HTS assay. Traditionally the quantitative components of these analyses are performed using Fluorescence, Luminescence, Absorbance, or Radiolabel techniques. These techniques provide rapid sample analysis but also contain inherent limitations, such as the occurrence of false positives, increased time for method development and complex sample preparation. High resolution accurate mass LC/MS coupled with multiplexing technology provides a selective and sensitive alternative to traditional methods without the need for labeled substrates and at rapid analytical speed. Samples were made to simulate an enzymatic screening assay and injected onto a C18 column (10x2.1mm, 3.5µm). Chromatographic separation was accomplished through a generic two step gradient using 0.05% trifluoroacetic acid in water and 0.05% trifluoroacetic acid in Acetonitrile. Mass spectrometry analysis was performed by heated electrospray in positive ion mode, and analyzed using generic MS conditions. Samples were injected to demonstrate reproducibility and calibration curves were analyzed to measure the dynamic range of the analytical method, along with the limits of detection and quantitation. The experiment data set containing 10,000+ sample injections provides results demonstrating CV’s of less than 20% for all sample replicates, linear signal response for all points across each calibration curve, and reasonable LOD and LOQ of 10nM and 25nm respectively, with an estimated analysis time of 2.25 hours per 384 well plate. This novel combination of hardware and software provides the unique capability to rapidly analyze large samples sets while retaining the ability to chromatographically remove unwanted. Through the implementation and integration of this technology, high quality MS data can now be collected at speeds capable of supporting the demands of large scale high throughput screening.
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