Identification of novel apoptotic markers based on hybridoma technology

Abstract

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.