 Cancer Protein Biomarkers Production
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Lee Makowski , Principal Investigator
Project Members: Diane J. Rodi, Frank Collart, Wen Zhang, Suneeta Mandava, Amina Aziz
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1G5M
Solution structure of the antiapoptotic protein bcl-2 |
1N9D
Solution structure of human prolactin |
Clinical Proteomics and Biomarker Discovery in Cancer Research
An effective platform for clinical biomarker discovery would revolutionize clinical oncology. Such a platform would enable detection of disease early, when it is curable. It would also provide a means of distinguishing quickly those patients who are responding to a given therapy from those who are not, as well as yielding perhaps the ultimate preventative tool capable of identifying those at risk for cancer and even prediciting their response to prevention interventions. Seldom during the span of a research career does one experience a technological breakthrough that, when applied in the world of clinical oncology, dramatically changes the lives of future patients. All indications are that we are at the vanguard of such an experience with programs to discover and validate cancer protein biomarkers. We can now imagine a day in the not too distant future when a routine visit to the clinic includes a health biospectral analysis - a simple blood test that analyzes thousands of proteins and genes that indicate susceptibility to cancer and detects cancerous cells at a sentinal stage.
The National Cancer Institute (NCI) has major initiatives to support the development and assessment of novel protein detection technologies. These technologies are designed to standardize proteomic technologies, reference reagents and standards, measurement protocols, instrumentation, and sample acquisition, preparation, and analysis platforms. A critical component needed to achieve the goals of these initiatives, will be the development and validation of proteomic reagents including peptides / proteins and antibodies. DOE has established unique capabilities and expertise in the production, qualification, and characterization of peptides and proteins. This experience currently supports the production of proteins that are used to develop crystal structures for the resolution of their 3-dimensional structures. Such high quality, well characterized, and readily available proteins and peptides will be a key resource in advancing proteomic technologies and capabilities and will be essential for developing highly specific antibody to proteins associated with cancer.
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NIST researchers are working to validate the mitochondrial DNA sequence measurement technology and increase the speed of the sequencing protocol, to provide improved methods for use in clinical applications.
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The goal of Argonne National Laboratory's NCI protein production program is the production, characterization, and distribution of peptides and proteins for use in the development and validation of clinical cancer proteomic technology platforms. Protein targets will be selected based on scientific rationale that considers the biological relevance of the targets in the cancer proteome. A candidate list of peptide and proteins for the proposed project will be selected by NCI in consultation with scientists in our group. We are responsible for the expression, purification, and characterization strategies for selected proteins and will provide the clones, proteins, data, datasheets, and protocols for NCI to be made publicly available for the scientific community.
Proteins will be expressed through a variety of strategies using E. coli expression systems. Protein targets will be chosen based on informatics-driven target categorization (e.g. size, localization signals, transmembrane-helices). The proteins will be generated from cDNA clones, RNA, or PCR-based gene synthesis. The protein constructs will then be purified and characterized by SDS-PAGE, mass spectrometry, circular dichroism, and light scattering. Each individual protein production process will be carefully tracked and detailed for successful expression strategies.
Project Milestones include:
- development of an initial target list of candidate human and murine proteins and peptides, including post-translationally modified targets
- analyze proposed targets and design protocols for each target
- carry out initial cloning, expression and solubility testing of each target protein
- scale-up successful clones to produce milligram quanitites of at least 100 proteins
- re-evaluate unsuccessful clones, redesign protocols then repeat cloning as necessary
Crystals of recombinant antibody |
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CSIRO is developing generic technologies and high affinity reagents for improved diagnosis and therapy of human disease. The design and production of multimeric recombinant antibodies as well as gene and protein array systems are being established to identify new biomarkers relevant to optimal human nutrition and health.
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See also : The National Cancer Institute's Clinical Proteomic Technologies for Cancer |
