Abstract
The two major methods of cancer therapy have been radiation and chemotherapy. Oftentimes these methods are successful, yet both treatments present enormous risk to healthy cells. Additionally, these treatments cause debilitating side effects and relapse is quite common. There is, however, a new direction for cancer therapy on the horizon called antibody-based immunotherapy. This new strategy utilizes the specific targeting property of antibodies to surround and remove tumor cells. The focus of my work was fusing a cytokine called GM-CSF to a single chain FvFc version of a recently FDA approved antibody called Rituxin. Rituxin is a chimeric antibody that recognizes a membrane bound phosphoprotein called CD20, which is expressed on the surface of all B-cells.The goal of this fusion protein design is to combine tumor specificity via the antibody with enhanced immune response activity via the cytokine. Such a design should endow the immune system with the ability to eliminate tumors from the body in a safe and efficient manner, thereby sparing millions of healthy cells and causing minimal side effects. A technique called Splice Overlap Extension PCR was the critical step in the gene design, which ultimately fused the two proteins together. Upon purification, sequencing, and transforming the DNA, the new gene was transfected into mammalian cells. Sadly, this is where the summer and my work ended. Future plans include purifying the protein, testing it for functionality, testing it in mouse trials, and ultimately putting it to use in human trials.
