Browsing by Subject "Hybridomas."

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Open Access
Identification of novel apoptotic markers based on hybridoma technology
(2004) Çıracı, Ceren
Apoptosis is a physiological cell death program that is characterized by morphological changes such as nuclear condensation, membrane blebbing, cell shrinkage, DNA cleavage and formation of apoptotic bodies. It is required for normal development and maintenance of cellular homeostasis and dysregulation of apoptosis leads to several developmental abnormalities, progression of degenerative diseases and cancer development. Elucidating the mechanisms underlying this form of cell death and development of molecular markers for its identification are of great importance to understand tumorigenesis. One way to generate molecular markers is based on monoclonal antibodyproduction. Utilization of such antibodies as apoptotic markers is strictly dependent on their molecular specificity and selectivity in terms of recognizing targeted proteins. We produced monoclonal antibodies by using apoptotic Huh-7 hepatocellular carcinoma cells as immunogen, and chose two of them, namely 9C11 and 6D5 for further characterization in the context of apoptosis.
Open Access
Investigation of novel tumor markers based on hybridoma technology
(2004) Çelikkaya, Hilal
Hybridoma technology is a highly specific technique utilized for the production of monoclonal antibodies. In cancer research, monoclonal antibodies are used as tumor markers for diagnosis of malignant tissue versus benign or normal, for differential diagnosis of tumor type, for pathological grading of tumor biopsy specimens, for detection of the antigens that are up- or down-regulated in tumor cells or in sera of cancer patients, and for detection of primary or metastatic lesions. Additionally, monoclonal antibodies are employed in predicting the course of the disease, in diagnostic approaches for carrying imaging reagents to tumors, and in therapy for targeting cytotoxic reagents to and triggering or blocking cell surface molecules. This study involved generation of 6D5 and 9C11 monoclonal antibodies against apoptosis induced hepatocellular carcinoma cell line HuH-7, followed by characterization experiments. 6D5 antibody recognized 5 different epitopes in a panel of 28 cell lines in Western blotting experiments. In immunohistochemistry studies, 6D5 demonstrated positive staining in cirrhotic and cancerous cells of liver cancer tissue samples. On the other hand, 9C11 antibody recognized a single band in the same panel of 28 cell lines but it was not immunoreactive in immunoperixodase studies of liver cancer tissue samples, under our experimental conditions.