Biotinylation Turnover Assay of the A33 Antigen




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Biotinylation Turnover Assay of the A33 Antigen Vivek Raman under the direction of Margaret Ackerman Massachusetts Institute of Technology Research Science Institute July 31 2007 Abstract Colon cancer is a leading cause of cancer death in the United States Current therapies such as chemotherapy will target and destroy both tumor cells and healthy cells Radioimmunotherapeutic treatment using the A33 antigen was found to have potential in speci cally targeting tumor cells The A33 antigen would be used in a two step radioimmunotherapeutic treatment involving administration of the A33 antibody which would bind to the A33 antigen expressed on the surface of tumor cells and administration of a toxin that will bind only to the A33 antibody and thus selectively destroy tumor cells In order to determine appropriateness of this antigen as a two step radioimmunotherapy target its turnover time in cultured colon epithelial cells was determined This was done by pulsed biotinylation of cell surface proteins followed by extraction of membrane proteins at di erent time points Western blots were then conducted and the bands of A33 were isolated The band density of the antigen at each time point was plotted yielding a graph of exponential decay from which the turnover time of the antigen was found to be signi cantly longer than the turnover time of bulk plasma membrane and other membrane proteins This indicates that the antigen is quite stable and thus has potential in the two step treatment against colon cancer 1 Introduction Colon cancer is the third most commonly diagnosed type of cancer and the second most common cause of cancer death in the United States 3 Before the tumor stage of colon cancer an adenomatous polyp forms in the colon which can be surgically removed preemptively preventing the spread of colorectal cancer ibid However once tumor cells begin to spread throughout the colon a di erent approach such as radioimmunotherapy is required A potentially useful target for the






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