Category: Assay Development and Screening
Conventional high throughput screening employs large decks of multiwell plates to test thousands of compounds against a particular target of interest. This method requires the use of specialized facilities and robotics, which make the cost of these screening campaigns a limiting factor in lead identification. Conversely, synthesizing high diversity libraries of compounds through simple chemistries on solid phase resin through a split-and-pool technique allows for rapid and inexpensive screening of large one-bead one-compound (OBOC) combinatorial libraries. Recently, a collaboration between the Paegel and Kodadek labs enabled further miniaturization of this platform through use of DNA-encoding. This allowed hit identification to transition from tandem mass-spectrometry deconvolution to DNA deep sequencing, which enabled use of smaller beads that are compatible with fluorescence-automated cell sorting (FACS) to streamline the screening process. In the most recent iteration of this screening platform, our lab employs weakly reactive electrophiles (WREs) as library diversity elements, providing the library with the ability to covalently modify a protein target of interest. This library has been screened against disease relevant 26S proteasome subunits and hits are being validated. Concurrently, new libraries incorporating additional WREs are being synthesized to increase the chemical space accessed by our OBOC DNA-encoded libraries.
Paige Dickson– Graduate Student, The Scripps Research Institute, Jupiter, Florida
The Scripps Research Institute
I am a third year graduate student who is interested in developing novel screening approaches for ligand discovery. My previous experience is in yeast display affinity maturation and protein chemistry. I am currently working towards the development of covalent screening using DNA encoding and flow cytometry based screening. Currently, our biological system of interest is the proteasome, which is a multi-protein complex responsible for most protein degradation within the cell.