Fast-Track proposals will be accepted.
Number of Anticipated Awards: 3-5
Budget (total costs, per award):
Phase I: $150,000 for 9 months;
Phase II: $1,000,000 for 2 years
It is strongly suggested that proposals adhere to the above budget amounts and project periods. Proposals with budgets exceeding the above amounts and project periods may not be funded.
The deadline for receipt of all contract proposals submitted in response to this solicitation has expired. It was: November 25, 2013 by 4:30 p.m. ET.
As phosphorylated proteins play a significant role in normal and abnormal cellular function, there is a critical need for the production of pure, analytically characterized phospho-proteins/polypeptides for use in assays designed to capture the phosphorylation signatures of different cancers. Phospho-protein/polypeptide standards can serve as reference controls for quantitative immunoassays, immunogens for generating phospho-specific antibodies, and can be utilized in kinase inhibitor screens as well as protein-protein interaction studies. Technologies for generating phospho-serine and –tyrosine proteins are currently progressing, whereas the production of phospho-threonine (pThr) proteins has been unsuccessful due to imprecise chemical, enzymatic, and recombinant techniques. For example, biological systems that generate pThr-reagents are lacking, and those that exist provide poor control over site-directed phosphorylation and the phosphorylation percentage at each site. Additionally, commercially available reagents often do not meet purity guidelines for specific analytic applications. Therefore, the development of an appropriate methodology for the production of high quality and analytically verified pThr-protein/polypeptide standards will be a valuable step toward the commercialization of assays that detect target proteins associated with disease and drug action. The identification of such phosphorylated biomarkers has significant potential for the development of safer and more effective targeted cancer therapies.
The primary goal of this contract topic is to develop innovative site-directed phospho-threonine protein synthesis technologies for the production of marketable pThr protein/polypeptide standards that enhance the analytic capacity of cancer assays. Technologies must demonstrate the reproducible generation of high quality, analytically verified phosphorylated threonine protein standards with 50 – 80% modification of the specified site. Protein standards listed below are of particular interest to the NCI; however, any phospho-protein target of clinical relevance to cancer may be developed. Peptide domains should contain 100 – 1000 amino acid residues that span the phospho-site of interest. A minimum of 30 amino acids in length may be acceptable, specifically if both a capture and detection antibody can bind to the peptide fragment.
Ultimately, the development of robust, quantitative, phospho-threonine specific assays will require:
|1||ATR (ataxia telangiectasia and Rad3-related protein)||pT1989|
|2||mTOR (mechanistic target of rapamycin||pT2446|
|3||Akt/PKB (v-akt murine thymoma viral oncogene homolog)||pT308|
|4||Chk2 (CHK2 checkpoint homolog)td>||pT68|
|5||53BP1 (tumor suppressor p53-binding protein)||pT543|
|6||XRCC1 (X-ray repair complementing defective repair in Chinese hamster cells 1)||pT519/T523/T488|