D.CSS_Attach1-SOW.pdf

Title of Project:

Statement of Need and Purpose:

Background Information and Objective:

Period of Performance:

General Requirements:

Specific Requirements:

Statement of Work (SOW)

Assay for the Detection of Lysobisphosphatidic Acid (LBPA)

The Therapeutics Development Branch Biology within the National Center for Advancing Translational Sciences (NCATS) needs assay development and optimization services for the detection of Lysobisphosphatidic Acid (LBPA).

Previous work in the field of lysosomal storage diseases has shown that lysobisphosphatidic acid (LBPA) is able to mobilize cholesterol in mammalian cells. The work conducted will target the LBPA concentration in cells. Specifically, the Contractor will search for compounds that elevate LBPA levels. The Contractor will use Niemann-Pick disease type C (NPC1) fibroblasts or neural stem cells for compound screening. The screen will be based on homogenous time resolved FRET technology. Compounds will be selected for those with the most efficacy to elevate LBPA with minimal toxicity. Those compounds will be confirmed using secondary assays.

Base Award - Task 1: Sixty (60) days.

In the event the options are exercised, the period of performance will be extended as follows:

Option Quantity 1 - Task 2: Sixty (60) days.

Option Quantity 2 - Task 3: Thirty (30) days.

Scope of Work:

Independently and not as an agent of the Government, the Contractor shall furnish all the necessary services, qualified personnel, material, equipment, and facilities, not otherwise provided by the Government as needed to perform the Statement of Work below:

Base Award - Task 1: Identification of Most Sensitive Donor-Acceptor Pair for HTRF Assay Development (Milestone 1): The contractor shall develop an assay for the detection of LBPA in cell-based assays using homogeneous time-resolved technology (HTRF) in high-throughput format (1536 wp or 384 wp).

The main task will be the assessment of best antibody pair or antibody-antigen pair. This includes: matrixing all pairs together, choosing top three pairs and running three-point concentration response, switching the donor and acceptor, determining the dynamic range of the selected pairs with a focus on sensitivity, lower limit of quantitation, and upper limit of detection.

Option Quantity 1 - Task 2: Optimization of Assay Window and Sensitivity (Milestone 2): The contractor shall select the obvious best antibody pair or antibody-antigen pair and begin optimizing donor and acceptor concentrations to obtain the highest assay window and sensitivity. If no one pair is obvious, the top two should be selected for further development. This includes a range of 4 or more concentrations for each donor and acceptor. Next, the contractor shall optimize volumes of the total reaction to mimic the cell-based assay in 384 or 1536 well plate format. 11-point concentration-response curves should be generated for the best pairs to determine the signal to background of all data points. The best optimization will be the one that has the highest sensitivity and the largest dynamic range. Media and cell lysates should be tested for detection of interference and LBPA in positive controls. Period of performance is sixty (60) days.

Option Quantity 2 - Task 3: Cell-based assay optimization (Milestone 3): The contractor shall evaluate the optimized reagents in 384 and 1536 well plates with cultured cells treated with positive control compounds.

Concentration response curves will be generated and full plate statistics will be obtained to determine assay suitability for high-throughput screening. Period of performance is thirty (30) days.

Level of Effort:

N/A

Government Responsibilities:

For Base Award Task 1 (Milestone 1): NCATS will provide the contractor with weekly/biweekly attendance on the team calls to review tasks and updates from the team. NCATS can provide the contractor with sources of antibodies, and literature regarding LBPA for NCATS’ applications.

For Option Quantity 1 - Task 2 (Milestone 2): NCATS will provide the contractor with weekly/biweekly attendance on the team calls to review tasks and updates from the team. NCATS can provide the contractor with sources of antibodies, and literature regarding LBPA for our applications. NCATS can provide positive and negative control cell lysates.

For Option Quantity 1 - Task 3 (Milestone 3): NCATS will provide the contractor with weekly/biweekly attendance on the team calls to review tasks and updates from the team. NCATS can provide the contractor with sources of antibodies, and literature regarding LBPA for our applications. NCATS can also provide cells and positive control compounds for testing.

Deliverables:

Milestone 1:

a. Milestone 1 Report: A report summarizing all data generated for all activities completed under Task 1 and delivered electronically to wei.zheng@nih.gov and kirill.gorshkov@nih.gov by the contract expiration date. The data shall be collected thoroughly with publication of the data in mind. Data sets shall be complete, and all omissions or augmentation of the data including individual data points, graphs, figures, etc. shall be reported in the detailed report.

Milestone 2:

a. Milestone 2 Report: A report summarizing all data generated for all activities completed under Task 2 and delivered electronically to wei.zheng@nih.gov and kirill.gorshkov@nih.gov by the contract expiration date. The data shall be collected thoroughly with publication of the data in mind. Data sets shall be complete, and all omissions or augmentation of the data including individual data points, graphs, figures,

b. 2,000 x 20μl tests of optimized reagents for LBPA assay: delivered by the contract expiration date to:

ATTN: KIRILL GORSHKOV

NCATS / NIH

9800 MEDICAL CENTER DR

BLDG B, RM 301

ROCKVILLE MD 20850

c. Protocol for HTRF assay: delivered electronically to wei.zheng@nih.gov and kirill.gorshkov@nih.gov by the contract expiration date.

mailto:wei.zheng@nih.gov mailto:kirill.gorshkov@nih.gov mailto:wei.zheng@nih.gov mailto:kirill.gorshkov@nih.gov mailto:wei.zheng@nih.gov mailto:kirill.gorshkov@nih.gov

Other Considerations:

Travel:

Key Personnel:

Information System Security Plan:

Data Rights:

Section 508:

Publications and Publicity:

Confidentiality of Information:

Milestone 3:

a. Milestone 3 Report: A report summarizing all data generated for all activities completed under Task 3 and delivered electronically to wei.zheng@nih.gov and kirill.gorshkov@nih.gov by the contract expiration date. The data shall be collected thoroughly with publication of the data in mind. Data sets shall be complete, and all omissions or augmentation of the data including individual data points, graphs, figures,

b. Protocol for HTRF assay in cell-based format: delivered electronically to wei.zheng@nih.gov and kirill.gorshkov@nih.gov by the contract expiration date.

Reporting Requirements:

In the event a Milestone is not completed within a month, a monthly progress report will be due for that month.

N/A

The Government retains unlimited rights in all data generated under this contract.

N/A

There shall be no publication of the data, results, findings, or conclusions generated under this contract without prior approval from the Government.

The contractor, including all its employees, consultants, and subcontractors, shall maintain confidentiality of all information generated under this project.

mailto:wei.zheng@nih.gov mailto:kirill.gorshkov@nih.gov mailto:wei.zheng@nih.gov mailto:kirill.gorshkov@nih.gov