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2005 Scientific Report

  • Text
  • Report
  • Institute
  • Mice
  • Proteins
  • Signaling
  • Protein
  • Michigan
  • Tumor
  • Molecular
  • Laboratory

External Collaborators

External Collaborators Philippe Chavrier, Institut Curie, Paris, France Jeff Frost, University of Houston, Texas Gregg Gundersen, Columbia University, New York George Prendergast, Lankenau Institute, Wynnewood, Pennsylvania Kathy Siminovitch, University of Toronto, Canada Recent Publications Alberts, A.S., H. Qin, H.S. Carr, and J.A. Frost. In press. PAK1 negatively regulates the activity of the Rho exchange factor NET1. Journal of Biological Chemistry. Eisenmann, K.M., J. Peng, B.J. Wallar, and A.S. Alberts. In press. Rho GTPase-formin pairs in cytoskeletal remodeling. In Signaling Networks in Cell Shape and Motility, London, U.K.: Novartis Foundation. Wen, Ying, Christina H. Eng, Jan Schmoranzer, Noemi Cabrera-Poch, Edward J.S. Morris, Michael Chen, Bradley J. Wallar, Arthur S. Alberts, and Gregg G. Gundersen. 2004. EB1 and APC bind to mDia to stabilize microtubules downstream of Rho and promote cell migration. Nature Cell Biology 6(9): 820–830. Left to right: Liu, Holman, DeWard, Chen, Peng, Collins, Alberts, Kitchen, Eisenmann 12

Laboratory of Antibody Technology Brian Cao, M.D. Dr. Cao obtained his M.D. from Beijing Medical University, People’s Republic of China, in 1986. On receiving a CDC fellowship award, he was a visiting scientist at the National Center for Infectious Diseases, Centers for Disease Control and Prevention (1991–1994). He next served as a postdoctoral fellow at Harvard (1994–1995) and at Yale (1995–1996). From 1996 to 1999, Dr. Cao was a Scientist Associate in charge of the Monoclonal Antibody Production Laboratory at the Advanced BioScience Laboratories–Basic Research Program at the National Cancer Institute–Frederick Cancer Research and Development Center, Maryland. Dr. Cao joined VARI as a Special Program Investigator in June 1999. Staff Ping Zhao, M.S. Tessa Grabinski, B.S. Laboratory Members Visiting Scientist Mei Guo, M.S. Students Yong-jun Jiao Xin Wang Jin Zhu Research Interests H epatocyte growth factor/scatter factor (HGF/SF) is a multifunctional heterodimeric protein produced by mesenchymal cells and is an effector of cells expressing the tyrosine kinase receptor Met. Met, the protein product of the c-met protooncogene, is from the same family as epidermal growth factor (EGF) receptors. The activation of Met by HGF/SF affects downstream signaling pathways (including other protein kinases) responsible for cellular differentiation, motility, proliferation, organogenesis, angiogenesis, and apoptosis. Aberrant expression of the Met-HGF/SF receptor-ligand complex—resulting either from mutations in the complex or in conjunction with mutations in other oncogenes—is associated with an invasive/metastatic phenotype in most solid human tumors. Met-HGF/SF and downstream kinases are therefore attractive targets for new anti-cancer agents for clinical diagnosis, prognosis, and treatment. The aberrant expression of the Met receptor kinase by two-thirds of localized prostate cancers, and apparently by all osseous metastases, suggests that Met provides a strong mechanism of selection for metastatic development. We have generated and characterized several anti-Met murine monoclonal antibodies (mAbs) that have high affinity for and specifically recognize Met extracellular domains in their native conformation. In collaborative studies, we are using two of these radiolabeled anti-Met mAbs, designated Met3 and Met5, to study mouse xenograft and orthotopic models of localized and metastatic prostate cancer via clinical nuclear imaging. Moreover, we will soon be testing these two radiolabeled mAbs on dog spontaneous prostate cancer and bone metastasis models. In collaboration with the Nanjing Medical University of China, we have initiated a project to construct a phage-display antibody fragment library. This technique involves the construction and use of animal/human, immunized/naïve Fab and scFv antibody gene repertoires by phage display. The ability to co-select antibodies and their genes allows the isolation of high-affinity, antigen-specific mAbs derived from either immunized animals or non-immunized humans. A number of procedures for selecting such antibodies from recombinant libraries have been described, and some useful antibodies have been produced with this approach. Over the past two years, we have closely followed the development of this technology for producing novel recombinant antibody-like molecules. We have constructed a human naïve Fab library with the diversity of 2 × 10 9 and have screened out some mAb fragments against tumor marker proteins. In particular, we have selected from this library and characterized one specific anti-Met Fab fragment, designated as Fab-Met-1, using a subtractive whole-cell panning approach (Figs. 1 and 2). We have also established the technology of a phage-display peptide library for mAb epitope 13

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