As part of the proteomics inititiative of NCI "cancer related proteins" are being produced and characterized which will be used to develop standards for proteomic efforts to identify protein biomarkers in serum. Our intitial target lists include a set of angiogenesis related proteins compiled as a result of our angiogenesis research, and a larger list compiled from an extensive literature search conducted by the Plasma Proteome Institute.

The development of the target lists include the following constraints:

• we will not produce cheap commercially available proteins
• we will incorporate integral membrane proteins into the list, but will delay attempts to produce them for at least 6 months and possibly longer
• we will attempt to express soluble domains of membrane proteins that might be of interest
• we will attempt individual domains of very large proteins (as well as full length proteins in some cases)
• if the full length clone is unavailable we may opt for gene synthesis
• intitial attempts will use prokaryotic vectors, so no post-translational modifications will be present in the expression products

Target lists of proteins currently being produced were screened from the following sources:

• Angiogenesis-related proteins - Our data demonstrates that a sequential upregulation of genes that establish and maintain polarity occurs during migration and morphogenesis of in vitro human endothelial cells undergoing tubulogenesis; some of which may well be effective as novel anti-angiogenic drug targets (Glesne et al, 2006).
• Extensive literature search conducted by the Plasma Proteome Institute has compiled a list of 1261 proteins which are believed to be differentially expressed in human cancer (Anderson, et al 2006).

Full length human or mouse clones have been purchased from Invitrogen or Harvard Institute of Proteomics. After screening for expression levels and solubility, milligram quantities will be produced of purified proteins. Initially all proteins will be produced in E. Coli vectors so they will not have post-translational modifications. Later expansion will include other vectors including rhodobacter vectors for integral membrane proteins, and eukaryotic vectors with the potential for more appropriate post-translational modifications.

We would be very interested in hearing about appropriate candidates for this effort, and then carrying out informatics analysis to determine the probability that our current vectors will be successful for their production. Appropriate candidates could then be integrated into our production pipeline. Investigators with potential targets can contact Diane Rodi.