Development of High Throughput Molecular Profiles for the Detection and Staging of Cancer

The discovery and identification of novel tumor associated antigens (TAAs) is critical for the advancement of accurate cancer diagnosis and staging. This Small Business Innovation Research Phase II project aims to continue development and application of anovel technology, Dual Immunostaining mediated Subtractive Cloning (DISC). The technology allows molecular cloning of genes that encode tumor cell surface proteins that are structurally or quantitatively different from those in normal cells. In Phase I,the feasibility of DISC was demonstrated. We have shown that rabbit polyclonal antibodies can be used as a complex probe to discover tumor associated cell surface antigens. Using the tumor cell line HeLa S3 and primary normal human fibroblast cells,polyclonal antibodies against these cell types were purified and effectively labeled using fluorescent dyes. A highly efficient method was established to identify cell surface antigens or unique epitopes, only expressed in tumor cells.In Phase II, DISC will be further developed and used to clone TAAs for human cancers of the breast, the prostate, and the lung. Tumor cell lines, tumor tissues, corresponding normal primary cells and normal tissues will be used to clone TAA genes. In thesame process, rabbit cell lines expressing these TAAs will be obtained. Rabbit monoclonal antibodies against these TAAs will be generated from these TAA expressing cell lines. Tumor specificity of the newly discovered TAAs and commercial potential of therabbit monoclonal antibodies will be evaluated by tumor tissue arrays.In this SBIR Phase II, Epitomics will discover and develop new TAAs and rabbit monoclonal antibodies that will enable 1) the rapid identification and characterization of novel tumor antigen targets and 2) commercial development of diagnostic kits.Epitomcis will supply the TAA genes and rabbit monoclonal antibodies to cancer researchers for clinical studies, commercialize them as research reagents and further develop some of the rabbit monoclonal antibodies into products for cancer staging anddiagnosis.