Characterization of an antigen-producing cell line hugk-14 at the molecular level

摘要

In order to evaluate its applicability as substrate for producing hepatitis B vaccine, a seed lot of human continuous cell line huGK-14 of hepatocellular carcinoma origin was established and investigated in its molecular genetic backgrounds. It was shown here that HBV DNA was integrated in eight different sites of the cellular DNA, in each of which HBV genome was rearranged, fragmented, and/or partly deleted. Complete HBV genome, as well as free HBV genome, was not detected in the cells nor in the culture medium. Clones of cDNA containing a complete coding frame for HBs antigen protein (type adr) were obtained. The cells were shown to be stable during more than 60 population doublings over the period of six months of cultivation in the mode of HBV integration and HBs mRNA expression. These results substantially fulfill the safety requirements for the seed lot to be used in vaccing production in terms of HBV genome integration and HBs gene expression.