Optimizing antibody production in batch hybridoma cell culture

Abstract

Optimizing productivity by hybridoma cells relies partly on developing suitable methods for screening and selection of high producing cultures and on understanding regulation of antibody production. In this study, the behavior of hybridoma cells in batch culture was investigated using flow cytometry, and a simple model for antibody production was used to explain production data obtained from these cultures. Surface antibody fluorescence values were found to closely follow the decreasing trend of specific antibody secretion rate over the course of several batch cultures. Therefore, for the hybridoma cell lines studied here (ATCC HB124 and TIB138), surface immunofluorescence levels can be used to select high producing cells as well as to monitor culture productivity. Surface and intracellular antibody fluorescence values were also found to be correlated for cells exhibiting a bimodal distribution with respect to intracellular antibody content. The population of cells containing lower levels of intracellular antibody was determined to secrete significantly less antibody than the population possessing high intracellular antibody concentrations. Factors which influence antibody production rates and possible strategies for optimizing monoclonal antibody yield are discussed.