The Monoclonal Antibody Facility provides Cancer Center and Northwestern University investigators access to the technology for the efficient creation of hybridoma cell lines and the production of monoclonal antibodies from these cell lines. These services include immunization of animals, somatic cell fusions, cloning and screening of hybridomas, subcloning and establishment of antibody producing cell lines, and production of active antibodies from hybridoma lines. In addition to providing these services, the facility provides consultation and training for NU investigators interested in establishing any of these activities in their own research laboratory or using monoclonal antibodies in their research. Surveys of researchers have led to projections that 91% of the usage of the facility will be in support of Cancer Center Members with peerreviewed funding. During the last grant period the facility has focused on providing its core services with higher efficiency and higher success rates, while increasing the user base. There are few examples of monoclonal antibodies relevant to cancer research that were generated by the facility during last grant period: 1. mAbs that recognize gp42. gp42 binds HLA Class II and is essential for EBV entry of B lymphocytes. These mAbs have been critical reagents in the characterization of gp42 domains essential for function. The mAbs are also being used is crystallography studies of gp42. 2. mAbs that recognize EIPR (E6-interacting p53 regulator), which is novel p53 interacting protein. This mAb is essential for ongoing study of novel E6 targets in epithelial cell biology and oncogenesis 3. mAbs directed towards a- methylacyl-CoA racemase (AMACR), which is highly expressed in prostate adenocarcinoma but not in benign prostate tissue 4. mAbs against angiopoietin-like 4 gene product (ANGPTL4 protein) that displays angiogenic activity. It was shown that ANGPTL4 is produced only in conventional renal carcinoma cells (conventional RCCs), and not in other benign or malignant renal legions, that makes it possible to use ANGPTL4 protein as a diagnostic marker for conventional RCC 5. mAbs that recognizes MAGE-D1, which is downregulated in subset of breast cancer cell lines. MAGE-D1 is likely a downstream target of BRCA2 and is required for BRCA2 mediated growth suppression. 6. mAbs recognizing Centrobin, which is also a BRCA2 binding protein. Centrobin is novel daughter centriole protein that is required for the centriole duplication. The anti-Centrobin monoclonal is currently being used to study the interaction BRCA2 and Centrobin and their functions.
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