Development and Implementation of an Antibody Array for Colon Cancer Screening
Colorectal cancer is the 4th most common cancer in the world. As with many cancers, early detection and screening are vital for reducing
morbidity and mortality associated with this disease. For colorectal cancer the existing modes of screening are not universally acceptable and
are not likely to be widely applied, particularly in developing countries. Therefore, we propose to produce an antibody array that will be applied to
discovery of blood plasma biomarkers for the early detection of colorectal cancer.
We will take a number of steps to ensure a positive outcome. We will utilize a multi-pronged approach to produce and select antibodies specific for a broad representation of plasma proteins including those of low abundance. This selection process will utilize conventional antibodies including the entire Development Hybridoma collection (NIH, University of Iowa) and monoclonals we are preparing from a mouse immunized with human sera. We will also screen a recombinant-antibody library with an innovative selection process. Approximately 5,000 plasma protein specific antibodies (conventional and recombinant) will be
utilized for the high-density array. This array will then be used to screen colorectal-cancer cases and controls. Quality control measures will be employed to ensure accurate screening and handling of large numbers of samples. Following screening, we will use biostatistical methods to evaluate and validate potential biomarkers. Finally, we will use biochemical methods to identify which antibodies bind potential biomarkers and then utilize these antibodies to identify their corresponding antigens.
This project is multidisciplinary in that it involves extensive antibody work and screening strategies to produce a viable array followed by careful and consistent application of the array in a case-control study and extensive biostatistical analysis to yield biomarkers that will be further investigated and identified using
immunohistological and Mass Spectrometry techniques.