D_Scope_of_Work_-_PSOL_attachment.docx

SCOPE OF WORK

GENERAL INFORMATION

Title of Project:

Generation and acquisition of 4000 custom 1536-Well NC Membrane Assay Plates.

Project Purpose or Objectives:

NCATS is in need of specific custom High Throughput Screening (HTS) Nitrocellulose (NC) Membrane Assay Plates for our 1536-well assays. This plate has been developed at NCATS for immunochemical and other array-based assays and is not commercially available. NCATS is currently using it for the development of reverse phase protein array (RPPA) assays but is also considering additional experimental applications. This requirement seeks a contractor to perform a custom-build of this plate and generate a large number of them for NCATS to use in multiple projects, thereby streamlining NCATS protocols.

The NC membrane plate prototype must meet certain user criteria, and must be compatible with all transport, storage, and robotic subsystems and components of the current in-house HTS systems. This NC membrane plate prototype is compatible with the 1536-well automation and instrumentation currently in use at NCATS.

Background Information:

The National Center for Advancing Translational Sciences (NCATS) at the National Institutes of Health (NIH) was officially established in fiscal year 2012 to transform the translational science process so that new treatments and cures for disease can be delivered to patients faster. NCATS, one of 27 Institutes and Centers (ICs) at NIH, strives to develop innovations to reduce, remove or bypass costly and time-consuming bottlenecks in the translational research pipeline in an effort to speed the delivery of new drugs, diagnostics and medical devices to patients.

Various programs and labs within NCATS have been given the directive to perform different types of high throughput screens to evaluate some biological activity and to potentially develop new therapeutics or therapeutic combinations in some cases or in others to determine the relative toxicity of chemical compounds under test. To do this, a biological experiment is performed in the presence of hundreds of thousands of chemical compounds at various concentrations, requiring that millions of samples be tested for each experiment. This requires that these experiments be run in a high density well plate, to both minimize the total reagent volume used per experiment and to facilitate lesser total plates be used to perform an entire assay. Typically, one experiment might consist of screening 500,000 compounds at 5 different total concentrations for a total of 2,500,000 samples. If this experiment is run using a 1536 well plate, meaning 1536 samples can be tested at a time, this requires nearly 1650 plates. The only way to accomplish handling this many well plates for an experiment is high throughput screening (HTS), which requires a high degree of robotic automation.

Over the past 10 years, NCATS has made a substantial investment in HTS systems; the two most active systems at the center are the Kalypsys Primary Screening system that was put into service in 2005 and the Tox21 Robotic Screening system that was put into service in 2007. Each of these systems requires the use of well plates, with the plate type to be used determined by the type of experiment being run. A white assay plate is typically used for a luminescence based experiment, while a black plate is used for a fluorescence based experiment. All plates used must be compatible with the HTS systems present at NCATS. Vendor’s plates must have been verified to be compatible with our systems.

This requirement is to obtain contractor-generated custom nitrocellulose (NC) membrane plates to be used on multiple robotic screening systems currently in use in NCATS in addition to the traditional wet lab research conducted for assay optimization and validation. Multiple labs within the institute have been given the directive to screen, identify, and distribute to the research community small molecule compounds, and data on their activities, to accelerate translation research of the completed human genome into advancements in human health. Components of NCATS include a repository of up to 500,000-1,000,000 chemical compounds, assay development capacity, robotic compound HTS screening, medicinal chemistry to transform ‘hits’ identified by HTS into workable chemical probes, chem-informatics capacity for data mining and virtual screening; and distribution capacity to efficiently disseminate the reagents to the biomedical research community.

Delivery Date:

Delivered in full to NCATS anticipated within 120 days after receipt of contract award

SCOPE OF WORK

General Requirements:

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 achieve the following specific requirements.

Project Requirements:

NCATS is in need of development, manufacture and delivery of 4,000 nitrocellulose membrane plates. They will be fabricated using three elements:

1. A 1536-well plate with no bottom element

2. Adhesive applied to well field top wall in the form of double sided sticky tape.

3. Nitrocellulose membrane (75mm X 110mm) applied to top surface of plate with adhesive providing bonding means to the well frame it is disposed upon.

The plate assay area must be capable of immobilizing a nitrocellulose membrane the same size as, or slightly larger, than the total well field on standard 1536-well assay plates using liquid or aerosol adhesive, double-sided adhesive tape, or other physical methods (sonic welding). The membrane is fully immobilized onto the plate frame top surface to give rigidity to the membrane and minimize distortion of the membrane material during spotting, blotting, washing and detection imaging development. The plate assay area must be a smooth top surface capable of providing a fixture to apply the nitrocellulose membrane securely to the top surface, and must be the same size or slightly larger than the total well field on standard 1536-well assay plates (75mm X 110mm).

The plate shape and dimensions are identical to normal high- or low-base 1536-well assays plates and will be compatible with plate storage and plate mobility (e.g. plate carousels, Kalypsys robotic arms), liquid handling (e.g. EDC Biosystems ATS), and reader/data acquisition instrumentation (e.g. Typhoon FLA 9500, Odyssey CLx, ViewLux, Envision) that are currently utilized at NCATS.

The nitrocellulose membrane plate must be capable of receiving small volumes of biological liquids in 384- and 1536-well array format (2.5nL – 1uL) using methods that include dispensing by acoustic droplet ejection or transfer by pin- or tip-based liquid handling instrumentation. The membrane blotting protocol conditions require immersion in fluids for extended periods of time. This aspect was tested thoroughly to show that the membrane remained fully bonded during the protocol steps.

The 1536-well plate must have the same footprint as a standard 96-well microplate to be adaptable to the existing UHTS robotic systems. The plate contains a membrane that is adhered to the frame of the 1536 wells, providing an open area under each well location for spotting of proteins and biomolecules onto the solid support.

1. Incubators, plate carousels, High Resolution and Kalypsys robotic arms, storage hotels

2. Liquid handling (e.g. EDC Biosystems ATS)

3. Reader/data acquisition instrumentation (e.g. Typhoon 9500, Odyssey CLx, ViewLux, Envision) currently utilized at NCATS

4. The current imaging method is placing the plates/membranes face down and using laser scanning instrumentation such as the GE Typhoon 9500 or Odyssey CLx. The preferred imaging method will involve automation-compatible readers such as the top-reading instruments ViewLux and Envision.

Essential features of plate size and dimensions follow:

a. The plates must conform to the size and dimensions of normal 384- and 1536-well assay plates currently in use to enable its compatibility with plate storage and handling, liquid handling, and reader/data acquisition currently utilized at NCATS

b. The plates should be barcoded using a method that is resistant to 24 hours of agitation in 1X TBS-T (50mM Tris-HCl pH 7.4, 150mM NaCl, 0.1% Tween-20)

c. The plate bottom support onto which the nitrocellulose membrane will be immobilized should be clear to enable visualization of spotting during transfer of liquid onto the plate using acoustic droplet ejection.

d. The nitrocellulose membrane should be pure nitrocellulose membrane and not “supported nitrocellulose membrane”

e. The nitrocellulose is cut to 75mm X 110mm and covers completely the well field.

f. The nitrocellulose membrane must have a maximum pore size of 0.45µm

g. Current source of nitrocellulose membrane: GE Amersham Protran Premium 0.45µm Nitrocellulose Membrane, 300mm x 4m 1 roll/pack.

h. The nitrocellulose membrane must be immobilized with liquid adhesive, double-sided adhesive tape or other physical methods that do not leach chemicals or background signal onto the nitrocellulose membrane

i. For physical immobilization methods, the nitrocellulose membrane must be mounted in a manner that withstands the inversion and dispensing process that occurs during acoustic droplet ejection by keeping the membrane stationary/in contact with the plate bottom support

j. The adhesive used must not fluoresce in far-red or near-infrared spectral ranges

k. The adhesive used must not dissolve or degrade nitrocellulose when used in the quantity needed for this application

l. The adhesive used must withstand a minimum of 24 hours in 1X TBS-T buffer with agitation (50mM Tris-HCl pH 7.4, 150mM NaCl, 0.1% Tween-20)

m. The nitrocellulose surface should contain the highest percentage of nitrocellulose obtainable to maximize binding capacity

n. The nitrocellulose surface should be as homogeneous across the plate as possible to minimize variability

o. The nitrocellulose surface used must not fluoresce in far-red or near-infrared spectral ranges

p. The nitrocellulose surface used must withstand a minimum of 24 hours in 1X TBS-T buffer with agitation (50mM Tris-HCl pH 7.4, 150mM NaCl, 0.1% Tween-20)

The COP material used to construct the 1536-well plate frame should be chemically inert, show low thermal deformation, be resistant to sterilization procedures, and be dimensionally stable. It should also minimize electrostatic effects. This may include the use of graphite-based polymers for electrical grounding or other electrostatic discharge methods. It should be of low autofluorescence black pigment to minimize background during detection and imaging of the NC membrane surface. The COP plastic will not be surface treated with RF microwave plasma.

Each 1536-well plate will be tracked in UHTS via its barcode label. The barcode shall be on a white label, one inch by 0.25 inches in dimension. There shall be a unique Code-128 barcode preprinted on the label. The label will be attached on the leading edge of the 1536-well plate (short edge) that faces the barcode readers in each UHTS subsystem.

The manufacturing process will occur in a clean room environment with special hoods and fixtures to enable dust free assembly of the NC membrane plate. Currently, the two key components of the product are brought together on the manufacturing line in a laminar hood (1536 COP frames and properly cut NC membrane in a stack separated by clean lint free sheet material).

Delivery Date / CONTRACT TYPE All deliverables shall be delivered to the Client in one (1) set no later than 120 days after receipt of an order. A Fixed-Price type of Purchase Order contract is anticipated to be awarded as a result of this requirement.

GOVERNMENT RESPONSIBILITIES

The Government will deliver all design requirements, protocols and test requirements needed for fabrication of custom plate to the successful contractor after receipt of any contract resulting from this solicitation.

DELIVERY OR DELIVERABLES

4,000 each 1536-Well NC Membrane Assay Plates, Barcoded, with plate lids

REPORTING REQUIREMENTS

All electronic concerns or progress reports should be delivered to NCATS via email.

TRAVEL

No travel cost shall be incurred by the contractor nor reimbursed by the Government.

KEY PERSONNEL

N/A

Confidentiality of Information:

Contractor will keep the details of the fabrication confidential.

DATA RIGHTS

NCATS shall have unlimited rights to and ownership of all deliverables provided under this contract, including reports, recommendations, briefings, work plans and all other deliverables. This includes the deliverables provided under the basic contract and any optional task deliverables exercised by the contracting officer. In addition, it includes any additional deliverables required by contract change. The definition of “unlimited rights” is contained in Federal Acquisition Regulation (FAR) 27.401, “Definitions.” FAR clause 52.227-14, “Rights in Data-General”, is hereby incorporated by reference and made a part of this contract/order.

Technical Evaluation / Selection Criteria: TBD