Studies on the growth and antibody production of a hybridoma cell line

• Main Author: Dempsey, Jonathan H.
• Published: University of Edinburgh 1994
• Subjects: 571.96
• Online Access: http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.649477

In this thesis a number of features of hybridoma cells have been investigated experimentally to obtain a better understanding of their growth and antibody secretion <I>in vitro</I>, with a view to improving the efficiency of antibody production. Initially the effect of immune derived mediators on the growth and secretion of hybridomas was explored as some of these mediators have been shown to play an important role in the growth and antibody secretion of B-cells <I>in vivo</I>. In particular interleukin-2 and interleukin-6 were studied and were shown, in some cases, to improve the rate of cell growth, but they had little or no effect on the level of antibody secretion. Using flow cytometry, the expression of antibody on the surface of a hybridoma cell was investigated. The presence of antibody on the cell surface was correlated with overall antibody secretion and to the stage of the cell cycle. It was shown that antibody that is in the process of being secreted from the cell can be detected and that it is related not only to the overall secretion of antibody but also to the stage of the cell cycle. Attempts to synchronize hybridoma cells in one phase of the cell cycle by well documented chemical means were unsuccessful, and a technique was developed to isolate a synchronous population using flow cytometry. A synchronous cell population was cultured and samples were analysed for cell surface antibody, stage of the cell cycle and antibody secretion at various points in the cell cycle.