D1 PD 061721.pdf

Version 11-30-2016-1 FAR Part 11—Describing Agency Needs

PURCHASE DESCRIPTION

1. Name of Item or Project Title: Purchase “HTRF® cAMP Gi Assay kit” and “HTRF® cAMP Gs Dynamic Assay kit” (Brand-name or equal).

2. Statement of Need and Purpose: The Tox21 group of National Center for Advancing

Translational Sciences (NCATS), Division of Preclinical Innovation (DPI) requires two reagent kits named “HTRF® cAMP Gi Assay kit” and “HTRF® cAMP Gs Dynamic Assay kit” (Brand-name or equal). These two reagent kits will use to screen Tox21 library collections (10,000 compounds) in a 1536-well high-throughput format. The purpose of the purchase is to screen Tox21 10K library to identify a group of G protein-coupled receptors (GPCR) for better biological response space coverage and toxicity prediction.

This project is one of the cross-partner projects in the Tox21 collaboration between

NCATS and NTP/EPA/FDA.

3. Background Information and Objective: The National Institutes of Health (NIH) is the nation’s leading medical research agency and the primary federal agency whose mission is to seek fundamental knowledge about the nature and behavior of living systems and the application of that knowledge to enhance health, lengthen life, and reduce illness and disability, conducting, supporting and making medical discoveries that improve people’s health and save lives.

The mission of the Tox21 group of NCATS at the National Institutes of Health (NIH) is to develop better toxicity assessment methods to quickly and efficiently test chemicals for their potential disrupt processes in the human body that may lead to negative health effects.

The Tox21 10K compound library has been screened against over 50 cell-based assays, in a quantitative high-throughput screening (qHTS) format, generating over 85 million data points, to date. Recently, we have initiated efforts to build computational models using these in vitro assay activity profiles to predict human toxicity including adverse drug effects. These assay data-based models achieved better than random, but <80% accuracy for most of these adverse effects. These results may be the consequence of insufficient coverage of the biological response space and lack of metabolic capacity in the cell lines used. Therefore, we initiated efforts to apply a more systematic and data driven approach to identify targets and pathways for better biological response space coverage and toxicity prediction. As surrogates of in vitro data, we incorporated some examples of known drug target annotation (DTA) information into these models, such as the protein/gene/pathway target of a drug or drug mechanism of action. Based on these results, we identified a list of targets/pathways not presently represented within the

Tox21 datasets that show promise of significantly improving the predictive power of the in vitro assay data on in vivo toxicity. A group of G protein-coupled receptors (GPCR) is one of these missing targets/pathways. “HTRF® cAMP Gi Assay kit” and “HTRF® cAMP

Gs Dynamic Assay kit” are the powerful tools to measure these receptors (Gs and Gi coupled receptors). We will build toxicity prediction models with these new assay data combined with the existing 10K data.

cAMP (cyclic adenosine 3’,5’-monophosphat) is one of the most important GPCR intracellular mediators of Gs and Gi coupled GPCRs. The “HTRF® cAMP Gi Assay kit” and “HTRF® cAMP Gs Dynamic Assay kit” developed by Cisbio uses HTRF® technology (homogeneous time-resolved fluorescence technology) to measure cAMP in a non-separation, high throughput format. The assay is specifically intended for the direct quantitative determination of cAMP. It is based on a competition between native cAMP produced by cells and cAMP labeled with the cryptate for binding to a d2 labeled antibody. The specific signal (i.e. energy transfer signal) is inversely proportional to the concentration of cAMP in the samples. This technology has successfully been used to screen several large libraries in 1536 well format, with high reproducibility and low false positives at NCATS. For example, the HTRF® TSHR assay was used to screen the

Tox21 10k library in 1536 well format with great success.

This purchase-needed project is for HTRF® cAMP Gi Assay kit and HTRF® cAMP Gs

Dynamic Assay kit to identify a group of G protein-coupled receptors (Gi and Gs) (Brand-name or equal) for better biological response space coverage and toxicity prediction.

These assay kits allow the measurement of agonist and antagonist effects on these receptors in different cell lines. The assay conditions have been optimized and validated in 1536-well high-throughput formats.

The assays are specifically intended for the direct quantitative determination of cAMP. It is based on a competition between native cAMP produced by cells and cAMP labeled with the cryptate for binding to a d2 labeled antibody. The specific signal (i.e. energy transfer signal) is inversely proportional to the concentration of cAMP in the samples.

The HTRF® technology brings major improvements in terms of handling. It is ideally suited for high-throughput applications with no washing steps involved. This type of assay is homogenous which can be miniaturized into 1536 well plate format for high throughput screening.

4. Generic Name of Product: competitive immunoassay intended to measure cyclic AMP

(cAMP) accumulation in cells.

4. Purchase Description:

• HTRF® cAMP Gi Assay kit, Part Number: 62AM9PEJ, Quantity: 10 ea.

• HTRF® cAMP Gs Dynamic Assay kit, Part Number: 62AM4PEJ, Quantity: 10 ea.

5. Salient characteristics: The salient physical, functional, or performance characteristics that “equal” products must meet are as follows:

• Gi Assay kit:

o The principle is based on HTRF® technology. It enables the direct compounds acting on Gs-coupled receptors in cells.

o Principles of the HTRF® technology: Anti-cAMP cryptate and d2-labeled cAMP are the two main components of the detection reagents. cAMP produced intracellularly is detected following cell lysis via competition between the intracellular cAMP and d2-labeled cAMP for the anti-cAMP antibody. An increase in intracellular cAMP leads to disruption of FRET signal, whereas a decrease in intracellular cAMP results in higher FRET signal.

o Shipping and Handling FOB destination.

• Gs Dynamic Assay kit:

o The principle is based on HTRF® technology. It enables the direct compounds acting on Gi-coupled receptors in cells.

o Principles of the HTRF® technology: Anti-cAMP cryptate and d2-labeled cAMP are the two main components of the detection reagents. cAMP produced intracellularly is detected following cell lysis via competition between the intracellular cAMP and d2-labeled cAMP for the anti-cAMP antibody. An increase in intracellular cAMP leads to disruption of FRET signal, whereas a decrease in intracellular cAMP results in higher FRET signal.

o Shipping and Handling FOB destination.

6. Quantity: 20 kits total (10ea - HTRF® cAMP Gi Assay kit, Part Number: 62AM9PEJ and

10ea - HTRF® cAMP Gs Dynamic Assay kit, Part Number: 62AM4PEJ.) (Brand-name or equal)

Delivery Date: one-time delivery is four (4) weeks after receipt of order (ARO). Delivery shall be made to the below address with FOB Destination.

National Center for Advancing Translational Sciences (NCATS) 9800 Medical Center Drive Rockville, MD 20850

7. Period of Performance: n/a. This is a one time delivery

8. Option(s): n/a