Project 3

Antibodies for Proteomics and Diagnostics (PI: S. Johnston)

We have developed a system for producing antibodies of exceptional quality in a high-throughput (htp) fashion. This system has been hardened so that we can meet the needs of the research community. We plan to continue this system as a service to meet the demands for research antibodies. We will expand the usefulness of these reagents by developing a system to screen them as potential biomarkers with an emphasis on heart disease. Therefore, in parallel we will add value to the antibody production by producing a set of reagents for use in human and rodent studies.

Specific Aim 1: Maintain and Improve the Antibody PGA Resource.
We will maintain the production of antibodies as requested by researchers, with a capacity of up to 2000/yr. The emphasis will be on the speed at which the antibodies can be made, and on antibodies for proteins that are not commercially available, have been difficult to make or for special applications. We will expand the service with large-scale production of specific antibodies and affinity purification of antibodies.

Specific Aim 2: Produce antibodies to the proteins expressed in the heart.
We will deploy the antibody production system to produce reagents for analyzing many of the proteins present in the heart. We will produce up to 200 affinity-purified antibodies per year in this effort, against rat and human proteins.

Specific Aim 3: Add value to the antibodies by putting in place a system to screen the heart antibodies as potential biomarkers.
Each antibody will be assessed for its 1) ability to detect a protein in the serum, 2) target being a stable marker in normal serum and 3) usefulness in assessing changes in the blood in response to ischemia in a rat model and human subjects.