Technical strengths

Discovery Technology

There is a great need to create novel approaches to drug discovery and development to optimize this critical process, as well as to create powerful pathway modulation tools for systems biology. The University of Pittsburgh Drug Discovery Institute (UPDDI) provides an environment in which novel technical approaches can be rapidly integrated into the drug discovery pipeline, along with existing methods.

Cell and Engineered Model Tissue Screening:

The staff of the UPDDI are highly experienced in cell analysis, from very high throughput whole well screens, to high throughput High Content Screening (HCS) and single cell multiplexed screens. The UPDDI maintains multiple HCS platforms, confocal and wide field, fixed endpoint and live cell, to automate the capture and analysis of fluorescence images of millions of individual cells exposed to thousands of compounds on a daily basis. Through optimal selection of appropriate cells, probes, antibodies, fluorescent protein fusions, biosensors and environmentally sensitive probes, the staff of the UPDDI can apply HCS technology to many drug target classes and phenotypic screens. HCS, whether confocal or wide field, is one of the most powerful tools available to investigate molecular mechanisms in the context of the cell and small experimental animal systems.

 

Zebrafish discovery:

The zebrafish embryo is an ideal animal system for in vivo HCS. We apply HCS to identify small molecules that modulate pathways in zebrafish embryos. The 1-day old embryo is approximately 1mm in diameter and can be easily arrayed into 96-well plates for screening. During the first day of development, embryos are transparent with most of the major organs present, thus enabling visualization of tissue formation during embryogenesis, and organ specific measurements.

 

Protein-protein interaction discovery:

We are combining novel computational biology and chemistry tools, including our in-house technology platform, AnchorQuery™, with very efficient, structure-directed MCR, X-ray crystallography, with fluorescence imaging methods to rapidly develop and validate panels of PPI's for research and drug discovery. AnchorQuery™ is a specialized pharmacophore search technology that brings interactive virtual screening of novel protein-protein inhibitors to the desktop. We are developing reversible PPI biosensors that will be incorporated into a variety of living and/or disease relevant cells to rapidly test for on- and off-target effects. This will permit the identification of cellular mechanisms of action (MOA's) and any off-target effects, including potential toxic liabilities.

 

High Content and High Throughput Screening:

The UPDDI Screening Facility, located on the ninth floor in the Biomedical Science Tower (BST), provides investigators with the ability to run High Throughput Screens in a variety of formats on hundreds of thousands of compounds. The facility is equipped with:

  • Liquid handling robots for compound dilutions, transfers and cherry picking
  • Multimode plate readers with luminescence, fluorescence, fluorescence polarization, time-resolved fluorescence, and absorbance detection capability
  • High Content imaging systems with automated plate handling
  • A cheminformatics Oracle database and a High Content Screening image storage and analysis database
  • Tissue culture hoods, incubators, plate sealing robots and all the other equipment needed to run high throughput screening campaigns on 1,000-200,000 compound libraries
  • QC procedures to ensure reproducibility