Identification of Novel Lung Tumor Proteins Using Tumor-Infiltrating B Cells

A multiple marker approach is a logical strategy to compensate for genotypic and phenotypic heterogeneity of lung cancer. Novel proteomic techniques are continually being applied to identify circulating proteins and expand the range of available lung cancer markers. We believe the antibody response to tumor antigens is a highly sensitive and specific indicator of the plasma proteome and provides a rational alternative to marker discovery. B cells that hone to the site of corresponding antigen (tumor-infiltrating B cells), are a self-selected tumor-specific subset of the patient's antibody repertoire. B cells isolated from fresh tumor and immortalized by Epstein-Barr Virus (EBV- TIB) are thus a concentrated and renewable source of tumor specific antibodies. Preliminary data shows how we have already identified four unique tumor antigens from limited screening of a phage-displayed tumor cell cDNA library with antibodies from cultured autologous EBV-TIB. This proposal describes our approach to library screening, protein identification, monoclonal EBV-TIB isolation, antibody validation and finally the use of antigen-specific human monoclonal antibodies to test for circulating proteins present in NSCLC patient, but not normal, plasma samples. In context, we hypothesize an assortment of antigen-specific human monoclonal antibodies to previously unknown tumor proteins can identify and dramatically expand the number of available cancer biomarkers measurable in the peripheral circulation. Commercially available multiplex assay platforms such as Luminex, designed for multiple biomarker measurement, will allow rapid translation from discovery to diagnostic stages of project development. Data from this innovative discovery-phase proposal will confirm proof of principle and be incorporated into a more comprehensive NIH proposal. University of Kentucky 1CY18105