Production of Monoclonal Antibodies against VACM-1/Cullin5
Dr. Maria Burnatowska-Hledin and Dr. Vicki Isola
Vasopressin – Activated Calcium Mobilizing receptor 1 (VACM-1) is a 780 amino acid long protein that is part of the E3-Ligase complex responsible for ubiquitin mediated protein degradation. VACM-1 has been shown to have severely decreased expression in cancer cell lines compared to normal cells. Until now it has been difficult to study VACM-1 in the cell using immunochemistry techniques (Western blotting, In-Cell Western, and immunostaining) to investigate cellular processes due to the limited availability of monoclonal antibodies against VACM-1. Whereas polyclonal antibodies can bind to multiple epitopes of a protein, monoclonal antibodies (mAbs) will bind to a specific epitope of a protein. The use of monoclonal antibodies conjugated with therapeutic drugs has become popular in cancer treatment. Monoclonal antibodies can be used to target the antigens that are uniquely expressed in cancer cells to deliver therapeutic drugs to cancer cells specifically or to stimulate the normal immune system pathways to help the body combat the cancer cells naturally. A new mAb against VACM-1 would be useful in providing insight into the role VACM-1 plays in cancer and how it might be used in cancer treatment. Prior to this study cell lineages that produce mAbs to VACM-1 were generated. The media from these cells (which should contain secreted antibody) were purified into fractions using column purification. Fractions from two of the cell lineages (1-10D and 2-F3) were found to contain antibodies. The aim of this study is to screen those fractions for biological activity.
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