Project Initiation (1 week)

Process

Principal investigaor (PI) identifies the most appropriate Core Facility for their specific project. PI may contact Thomas “TC” Chung for help in indentifying the most appropriate core facility. PI contacts Director of the selected Core Facility (CPCCG team). Alternatively, PI could contact Shakeela Dad (CPCCG Outreach Coordinator) for help in directing to the most appropriate Core Facility. PI and Core director organize an initial meeting (within 1 week). Single-stage projects are normally attended by Core Facility director For multistage projects where a transition of the project between the facilities is expected the presence of several/all CPCCG directors may be required PI outlines the biology of the system and the goals of the project Core director provides preliminary evaluation of the existing approaches Initial plan of action is discussed

Assay Development (~1-4 months)

Process. Technology identification, design and development (0-3 months).
Assay development director performs search and analysis of the technical and scientific literature: To explore the potential of using an assay from similar systems To identify other possible approaches To verify an initial hypothesis on applicability of a novel detection technology Assay development director and the PI arrange another meeting, normally within 1 week of the first one: To discuss additional strategies for the project To adjust and accept the plan of action and to identify further steps To outline investigators’ responsibilities and timelines CPCCG Core personnel performs the experiments: To test the selected detection technology To optimizes detection technology for the need of the particular assay To defines limitations of the detection system
Process. Assay Implementation (1-2 months).
Responsibilities - CPCCG	Responsibilities – PI Lab
Guide PI Lab on appropriate assay formats and protocols Develop primary HTS assays Review data generated by PI Provide an SOP for the assay	Ensure that proposed assay is consistent with the biology Provide specialty reagent for the assays Provides an SOP for the assay

If PIs would have their assay developed by own lab personnel, the assay development or high content websites could provide some useful links.

Screening (~1-2 months. Allow more time if using high-content screening)

Process
PI lab and the Chemical library and Screening (CLS) core conducts a pilot screen using a 1280 compound library (LOPAC from Sigma), which is on four (4) 384 well plates. PI lab and CLS core evaluates the pilot screen data, if this data meets the HTS criteria, a second pilot screen of nine (9) 384 well plates is performed using the 2000 compound Spectrum collection and 450 compound NIH Clinical Collection. PI lab and CLS core evaluates the second pilot screen data, if this data meets the HTS criteria, then precede to a 20 plate screen. The second pilot screen can be by-passed Screening can be stopped after pilot phase PI lab and CLS core evaluates the 20 plate screening data. PI lab accepts the data then further modifications to the SOP are made accordingly. CLS core will now scale up the 20 plate screening to 30-100 plates per run. CLS core enters the screen to a queue (normally the screening will take place shortly after the smaller screens are completed).
Process. Assay Implementation (1-2 months).
Responsibilities – CPCCG	Responsibilities – PI Lab
Manage and supply compounds for the screens Load all data into CBIS (Burnham’s biological database) Evaluate progress of screening workflow Provide automation expertise and guidance	Provide dedicated personnel for running the screen Write a final SOP for large scale screening (if undertaken) Provide all consumables and reagents to run the screen

For an additional fee, CPCCG will perform the screen without assistance from the PI Lab.

Follow-up

Process
CLS core will create a report summarizing the HTS campaign This report includes all calculated data and a summary of all hits PI Lab and CLS core will determine which compounds to cherry pick Compounds can only be supplied in 100% DMSO for: LOPAC Spectrum NIH Clinical Collection Only enough compound can be resupplied for duplicate, single concentration testing (not for dose response testing) PI lab and CLS core will reconfirm hits. PI lab reorders dry powder compounds. PI lab performs dose response and secondary assay experiments. Assay development or HCS facilities can perform these studies if requested. Additional assays are offered to test compounds for selectivity, specificity, solubility, and cytotoxicity.
Responsibilities – CPCCG	Responsibilities – PI Lab
Provide HTS campaign report Provide minimal cherry picked compounds Assist with hit reconfirmation Perform dose response and secondary assays Perform panel studies for selected compounds Provide hit confirmation summary	Assist with cherry picking and hit reconfirmation. Reorder dry powder compounds. Communicate progress to screening facility.

If you have any questions please contact:
Dr. Eduard Sergienko (Director, Assay Development)
Dr. Susanne Heynen-Genel (Director, High Content Assays)
Dr. Steve Vasile (Director, Compounds and HTS Facility)