Research Project
9202440
Project Summary Antibodies can facilitate the study of associations between different proteins in the cell using a technique known as immunoprecipitation (IP). This approach can reveal differences in protein associations present in healthy vs. diseased tissues, provide targets for diagnostics and therapeutics, and reveal protein--protein interfaces that could be druggable targets. It is widely recognized that a lack of high quality antibodies against human proteins is negatively impacting biomedical science. CDI Laboratories, Inc., has developed a pipeline that employs the largest content full--length human protein array to generate quantifiably monospecific mouse monoclonal antibodies (mAbs), and has generated a large number of mAbs against human transcription factors (TFs). The PI of this project has developed methods that enable and optimize the capture of endogenous protein complexes by IP in conjunction with mass spectrometry analysis. In Phase I of this STTR project, Aim 1 will characterize anti--human TF mAbs for their ability to IP endogenous cancer--related TF protein complexes. This will be accomplished initially using human tissue culture cell lines to establish quantitative functional metrics of the mAbs, and then applied to protein complexes isolated from patient-?derived cancers. In Aim 2, cancer--related TFs will be used as bait for IP of native complexes, and novel mAbs will be generated by immunization with the endogenous TF protein complexes. The target specificity of the mAbs will be determined using the human proteome array, yielding a catalog of new, high quality mAbs with which to study protein-?protein interactions within cancer cells. The long term goal of the project, which will be expanded and achieved in Phase II, is the production of larger numbers of high quality mAbs against native human proteins that are normally found as constituents of complexes in healthy and cancerous cells.
