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.
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