(Fast-Track proposals will be accepted.)
Number of anticipated awards: 4
Budget (total costs): Phase I: $200,000 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 13, 2012 by 5 p.m. EST.
Anti-peptide capture reagents can be used to identify and quantify proteins containing a target peptide sequence with a number of applications in biological research and bioassay development. For instance, they are used routinely in techniques such as immunoprecipitation, Western blot, and immunohistological identification and protein localization. A recently-developed application of such reagents is Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA), which utilizes antibodies to enrich peptides from complex matrices for quantitation of proteins by stable isotope dilution mass spectrometry. SISCAPA has the potential for simultaneous quantification of multiple targets from a given sample. There is growing interest in the development of such multiplex protein assays, including large-scale biomarker candidate verification studies and analyses of targeted biological pathways. Concurrent quantification of multiple protein analytes is highly desirable in these applications to minimize sample requirements, handling, and assay costs per analyte, while maximizing throughput.
The goal of this project is to develop new technologies that generate reproducible, well-characterized anti-peptide capture reagents for use in affinity-enriched proteomic studies for the cancer research community. An important characteristic of the desired reagents is the ability to immunoprecipitate their target peptide with high affinity. These reagents should be comparable or superior to ELISA-based antibody technologies in terms of specificity, affinity, and sensitivity and be reproducibly generated in a cost-effective and efficient (e.g. renewable) manner. Currently, mice are often used to generate monoclonal antibodies or alternative capture reagents, but peptides do not always elicit a potent immune response, which can result in low yield of antibodies to effectively immunoprecipitate the target peptides. The desired technology will likely be one that produces a strong immune response to peptide antigens, and may include the use of species other than mice for the generation of antibodies, since other species may have more diverse epitope recognition and improved immune response to small-size epitopes.
The development of these affinity capture reagents will be done in coordination with NCI's Clinical Proteomic Technologies for Cancer (CPTC) group. A list of proteins or proteotypic peptides derived from cancer biomarker candidates may be requested from CPTC. Furthermore, these capture reagents must be made available as a resource to the scientific community. The suggested choices of performance platforms that the affinity reagents must be compared to include mass spectrometry-based quantitative assays, immunoprecipitation, ELISA-based assays, Western blot, and immunohistochemistry. In addition, other considerations should include sensitivity/specificity/affinity information for the reagents, and method comparison with gold standard practices, precision, and LOD/LOQ.