SOW.doc

STATEMENT OF WORK

(2-17-2019) Requested Vendor Name: Molecular Devices

1.0

PURCHASE ORDER TITLE

Purchase of a Refurbished FLIPR Tetra to replace NIH # 01299939 2.0

BACKGROUND

The NIDDK and all its laboratories utilize the current FLIPR Tetra system manufactured by Molecular Devices. The current most frequent users are the LBC sections, i.e. MRS and MSS. The current FLIPR system was purchased in 2004 for a cost of $329,000. The 15-year-old unit has had several major failures in the recent past. The research studies the LBS and MSS conduct require the measurement of membrane receptor activity. The reaction that is generated must be measured instantaneously and requires very sensitive detectors. Very sensitive cameras are used along with a robotic injector system to facilitate the instant read from time zero. Reactions are followed in time, simultaneously in each well of a 96-well plate, by sensitive measurements obtained from the detector camera.

Ligand interactions with receptors and the cellular response to ligands interactions with receptors cannot be measured effectively without instant read technology.

The LBC laboratories require this data for all publications demonstrating the effects of ligands on receptor binding to both active and allosteric sites and the intracellular cascades that are caused by the activation (agonism) or deactivation (antagonism) of the receptor.

This SOW is for the purchase of a Fully factory refurbished FLIPR tetra at a substantially reduced cost with an ICCD camera for both fluorescence and luminescence to replace our current FLIPR Tetra. The unit will be refurbished by the original manufacture (Molecular Devices) and have full original equipment manufacturer one-year warrantee.

The following is a list of recent publications that required the data generated from the FLIPR for their publication in scientific Journals.

MRS section Gao, Z.G., Inoue, A., Jacobson, K.A. On the G protein-coupling selectivity of the native A2B adenosine receptor. Biochem. Pharmacol., 2018, 151:201-213.

Toti, K.S., Jain, S., Ciancetta, A., Balasubramanian, R., Charkaborty, S., Surujdin, R., Shi, Z.D., Jacobson, K.A. Pyrimidine nucleotides containing a (S)-methanocarba ring as P2Y6 receptor agonists. Med. Chem. Comm., 2017, 8:1897-1908.

Gao, Z.G., Jacobson, K.A. On the selectivity of the Gαq inhibitor UBO-QIC: A comparison with the Gαi inhibitor pertussis toxin. Biochem. Pharmacol., 2016, 107:59-66

Gao, Z.G., Balasubramanian, R., Kiselev, E., Wei, Q., Jacobson, K.A. Probing biased/partial agonism at the G protein-coupled A2B adenosine receptor. Biochem. Pharmacol., 2014, 90:297-306.

Costanzi, S., Kumar, T.S., Balasubramanian, R., Harden, T.K., Jacobson, K.A. Virtual screening leads to the discovery of novel non-nucleotide P2Y1 receptor antagonists. Bioorg. Med. Chem., 2012, 20:5254-5261.

Wei, Q., Costanzi, S., Liu, Q.Z., Gao, Z.G., Jacobson, K.A. Activation of the P2Y1 receptor induces apoptosis and inhibits proliferation of prostate cancer cells. Biochem. Pharmacol., 2011, 82:418-425.

MSS section Pydi SP, Jain S, Tung W, Cui Y, Zhu L, Jain S, Abel BS, Skarulis MC, Liu J, Caron MG, Gavrilova O, Finkel T, Wess J. Adipocyte -arrestin-2 is essential for maintaining whole body glucose and energy homeostasis. Nat Comm, accepted in principle Rossi M, Zhu L, McMillin SM, Pydi SP, Cu Y, Lee RJ, Kaneto H, Birnbaum MJ, Ma Y,Rotman Y, Liu J, Cyphert TJ, Finkel T, McGuinness OP, Wess J. Hepatic Gi signaling as a critical novel regulator of whole body glucose homeostasis. J Clin Invest 128, 746-759, 2018.

Zhu L, Rossi M, Cui Y, Lee RJ, Sakamoto W, Perry NA, Urs NM, Caron MG, Gurevich VV, Godlewski G, Kunos G, Chen M, Chen W, Wess J. Hepatic -arrestin-2 is essential for maintaining euglycemia. J Clin Invest 127, 2941-2945, 2017.

Hu J, Stern M, Gimenez LE, Wanka L, Zhu L, Rossi M, Meister J, Inoue A, Beck-Sickinger AG, Gurevich VV, Wess J. A G protein-biased designer G protein-coupled receptor useful for studying the physiological relevance of Gq/11-dependent signaling pathways. J Biol Chem 291, 7809-20, 2016 (paper of the week).

Rossi M, Ruiz de Azua I, Barella LF, Sakamoto W, Zhu L, Cui Y, Lu H, Rebholz H, Matschinsky FM, Doliba NM, Butcher AJ, Tobin AB, Wess J. CK2 acts as a potent negative regulator of receptor-mediated insulin release in vitro and in vivo. Proc Natl Acad Sci USA 112, E6818-24, 2015.

Hu J, Hu K, Liu T, Stern MK, Mistry R, Challiss RA, Costanzi S, Wess J. Novel structural and functional insights into M3 muscarinic receptor dimer/oligomer formation. J Biol Chem 288, 34777-90, 2013.

Li JH, Jain S, McMillin SM, Cui Y, Gautam D, Sakamoto W, Lu H, Jou W, McGuinness OP, Gavrilova O, Wess J. A novel experimental strategy to assess the metabolic effects of selective activation of a Gq-coupled receptor in hepatocytes in vivo. Endocrinology 154, 3539-51, 2013.

Nakajima KI, Wess J. Design and functional characterization of a novel, arrestin- biased designer G protein-coupled receptor. Mol Pharmacol 82, 575-82, 2012.

Designer Receptors Exclusively Activated by Designer Drugs. Part of the Neuromethods book series (NM, volume 108) Chapter 2 Design and Analysis of an Arrestin-Biased DREADD; Ken-ichiro Nakajima, Luis E. Diaz Gimenez, Vsevolod V. Gurevich, and Jürgen Wess 3.0

OBJECTIVE

The objective is to obtain a fully functional instant read robotic microtiter plate reader capable of measuring both fluorescence and luminescence that auto-injects simultaneously and reads all wells continuously for the duration of the reaction. This unit will be for the use of all NIDDK as well as LBC staff.

Photographic instant read, and robotic injection technology are required for any research that measures the effects of endogenous molecules, drugs, and drug like compounds, on cellular response to receptor activation (agonism) or deactivation (antagonism). Our research uses both fluorescence detection and luminescence detection.

Measurements of membrane potential, signaling cascades, and ion uptake (Ca2+) are via fluorescent excitement and detection. The reaction that is generated must be measured instantaneously and requires very sensitive detectors. Instant read from time zero is an absolute requirement. Reactions are followed in time by sensitive measurements with the detector camera.

Ligand interactions with receptors and the internal cellular response to ligands interactions with receptors cannot be measured effectively without instant read technology.

In addition, the research into signal transduction requires the measurement of interaction between both fluorescent and luminescent tagged moieties BRET; (BREt allows for the monitoring of the physical interactions between two proteins, one of which is fused to a bioluminescent donor moiety and the other one to a fluorescent acceptor moiety allowing for measurements of resonance energy transfer between these two moieties) 5.0

TASKS AREAS

Independently, and not as an agent of the Government, the contractor shall perform all of the detailed Task Areas listed below Work to be completed by vendor:

· Installation including all components, detectors, cameras, computer and software.

· Inside delivery (Bldg. 8, Room 120)

· Unpacking of boxes and disposal of shipping and packing materials after installation

· On-site training one day for 4 users. NIH requires additional training days if the training cannot be completed in one day.

6.0

PURCHASE ORDER TYPE

This purchase order will be a firm fixed price, with payment terms per the vendor allowance based on whether it is a small business (net 15), or a large (net 30).

7.0

PERIOD OF PERFORMANCE

Delivery before June 1, 2019 and one-year service from installation date.

8.0

INSPECTION AND ACCEPTANCE

The Project Officer Jeanne Kaufman, or the Project Officer Supervisor Kenneth Jacobson will perform inspection and acceptance of materials and services to be provided. Inspection and acceptance will be done AFTER All materials are unpacked, installed, and test run for operation. Receiving will be completed no later than 7 days following date of Delivery and DOES NOT constitute acceptance and Delivery. Acceptance and Delivery will be complete once the signed packing slip is provided to the Vendor.

Inspection and acceptance will be performed at the place of installation - National Institutes of Health, NIDDK, 9000 Rockville Pike, Maryland 20892.

9.0

PROJECT OFFICER

The following Project Officer(s) will represent the Government for the purpose of this Purchase Order:

Project Officer: Jeanne Kaufman

Alternate Project Officer: Kenneth Jacobson

The Project Officer is responsible for: (1) monitoring the Contractor's technical progress, including the surveillance and assessment of performance and recommending to the Contracting Officer changes in requirements; (2) interpreting the statement of work and any other technical performance requirements; (3) performing technical evaluation as required; (4) performing technical inspections and acceptances required by this Purchase Order; and (5) assisting in the resolution of technical problems encountered during performance.

The Contracting Officer is the only person with authority to act as agent of the Government under this Purchase Order. Only the Contracting Officer has authority to: (1) direct or negotiate any changes in the statement of work; (2) modify or extend the period of performance; (3) change the delivery schedule; (4) authorize reimbursement to the Contractor any costs incurred during the performance of this Purchase Order; or (5) otherwise change any terms and conditions of this Purchase Order.

PAGE