With access to state-of-the-art robotic equipment our High-Throughput(HT) Biology Core can quickly screen and identify putative small molecule inhibitors or critical host cell factors required for replication using BSL2, BSL3, or Select Agent pathogens. We will work with you to develop customized high-throughput assays to meet your research needs. Once an assay has been validated for screening our team can provide primary, dose-response, confirmation, and counter-screening for both small molecules and siRNA using in-house libraries. Our experienced staff will review and validate screening results and assist with data analysis to help you select potential leads with confidence. We are also able to provide follow-up guidance for mechanism of action studies to confirm target molecules as a viable probe to initiate a hit-to-lead discovery program for a potential new therapeutic or biological research tool.
In addition to focused drug discovery services, our team provides support for most molecular biology needs in a HT format including RT-PCR, real-time RT-PCR and more!
Enamine library (10,000 compounds)
Selected compound collection available from Enamine. Excludes non-druggable, promiscuous, reactive, and other undesirable structure types as determined using traditional (e.g., [loosened] Lipinski rules, undesirable functionality filters) criteria. The assembled collection consists of over 1,000 Murcko scaffolds, each present in a cluster size of 10-15 members in order to provide some minimal SAR data directly from the primary screen.
Silencer® Mouse Druggable Genome siRNA Library V3
A total of 19,344 unique siRNAs targeting 6,448 genes; 3 siRNAs per gene.54 plates with 352 siRNAs each,3 plates with 112 siRNAs each.
Silencer® Select Human Druggable Genome siRNA Library V4, 384 Well Plates
A total of 27,093 unique siRNAs (0.25 nmol) targeting transcripts from each of 9,031 human genes. 75 plates with 352 siRNAs each. 3 plates with 232 siRNAs each.
ABI 7900HT Real time PCR
96-, 384-well, and TaqMan® Low Density Array formats
Robotic arm for batch processing
ABI PCR Verti 96
0.1 mL volume
Capillary based protein RNA/DNA analysis
8-channel medium throughput automatic liquid processing
96-channel pipette heads for drugging 96-well format
Biotek Synergy 4
High-performance filter-based and monochromator-based detection
Biotek EL406 Microtiter Plate Washer
Washes 96- or 384-well plates
We will work with you to develop and provide to most cost-effective, full-service assays designed to meet your research needs. Per sample and per plate options are available for most services. Please contact us for the most up-to-date availability and pricing.
Getting a cell-based assay established for high-throughput screening (HTS) can be challenging. Some suggestions for moving forward on your assay are provided below. We typically use a two-step approach to validate cell-based assays for viruses or viral reporters for use in HTS platform:
Step 1-Determine the culture conditions to yield optimal dynamic range. Automation of a cell-based assay in a 384-well plate format requires obtaining the optimal cell and virus culturing conditions to achieve the optimal dynamic range. We recommend starting with a 96-well format before moving to 384 to titrate the multiplicity of infection (MOI) and number of cells. Generally, an HT screen will be effective if the signal generated by the cells has a difference of five times greater than that generated by the cell plus the reporter. The greater the difference between the reporter and the cells alone, the more sensitive the assay; and the more likely the assay will be to capture compounds that have lower potency. In addition to MOI and cell numbers the following parameters should be considered:
- optimal conditions for cell/pathogen growth
- kinetics of infection and optimal time point to endpoint readout
- effect of DMSO on the above parameters
- control drug
Step 2-Determine assay robustness. Now that you have established the best possible conditions to give you the optimal signal to background, focus on the robustness of your assay. Assay robustness is defined as the consistency of signal for each of the experimental conditions, and the dynamic range. For example, prepare a 384-well plates with each of the following: cells alone, virus plus cells, cells + DMSO, or virus + control drug. Your assay is considered successful when the variation for each condition is less than 10% from well to well. To confirm the consistency of the dynamic range, 384-well plates are prepared containing uninfected cells and infected cells, and the signal levels determined. The experimental results provide data on the variability of the two conditions. These data are then subjected to a Z-value statistic to determine the number of replicates needed for each compound. For a large screen, generally greater than 50,000 compounds, the Z-value should be >0.5 indicating that each compound needs only to be tested once. These experiments are repeated at least three times to prove consistency of the assay.