Generation of a Monoclonal Antibody Specifically Reacting with Undifferentiated Liver Progenitor Cells

Abstract

An adult rat liver progenitor cell line Lig-8 was established. In the induction by sodium butyrate, these cells were shown to be able to differentiate into both hepatocytes and bile duct cells expressing albumin and cytokeratin-19, the markers of respective cell types. In order to generate Lig-8 specific antibody for further studies, we produced a monoclonal antibody using the whole Lig-8 cells as immunogen. The yielded monoclonal antibody, named Ligab, belongs to IgG subclass G1 and κ light chain. It specifically stained on Lig-8 cells in the cytoplasm but not on a rat hepatoma cell line H4IIE. Its immunoreaction against Lig-8 cell lysate on dot blots diminished as the concentration of sodium dodecyl sulfate (SDS) in the lysate increased to 2%, a level in the sample buffer of standard SDS-polyacrylamide gel electrophoresis (PAGE). Not surprisingly, Ligab failed to detect its reacting antigen in Lig-8 cell lysate by standard SDS-PAGE-based immunoblotting. It could detect this antigen only by native PAGE-based immunoblotting. These characteristics suggested that the antigenic epitope for Ligab is likely a molecular structure instead of a peptide sequence. More interestingly, expression of Ligab-reacting antigen in Lig-8 cells declined as the cells were induced to differentiate by sodium butyrate. This antigen is very likely a differentiation-related marker for these cells, and this monoclonal antibody may help study the molecular mechanisms of liver progenitor cell differentiation.